Aequorin: A photoprotein isolated from the bioluminescent jellyfish Aequorea. It emits visible light by an intramolecular reaction when a trace amount of calcium ion is added. The light-emitting moiety in the bioluminescence reaction is believed to be 2-amino-3-benzyl-5-(p-hydroxyphenyl)pyrazine (AF-350).Scyphozoa: The class of true jellyfish, in the phylum CNIDARIA. They are mostly free-swimming marine organisms that go through five stages in their life cycle and exhibit two body forms: polyp and medusa.Luminescent Measurements: Techniques used for determining the values of photometric parameters of light resulting from LUMINESCENCE.Ferrets: Semidomesticated variety of European polecat much used for hunting RODENTS and/or RABBITS and as a laboratory animal. It is in the subfamily Mustelinae, family MUSTELIDAE.Calcium: A basic element found in nearly all organized tissues. It is a member of the alkaline earth family of metals with the atomic symbol Ca, atomic number 20, and atomic weight 40. Calcium is the most abundant mineral in the body and combines with phosphorus to form calcium phosphate in the bones and teeth. It is essential for the normal functioning of nerves and muscles and plays a role in blood coagulation (as factor IV) and in many enzymatic processes.Arachnida: A class of Arthropoda that includes SPIDERS; TICKS; MITES; and SCORPIONS.Luminescence: Emission of LIGHT when ELECTRONS return to the electronic ground state from an excited state and lose the energy as PHOTONS. It is sometimes called cool light in contrast to INCANDESCENCE. LUMINESCENT MEASUREMENTS take advantage of this type of light emitted from LUMINESCENT AGENTS.Luminescent Proteins: Proteins which are involved in the phenomenon of light emission in living systems. Included are the "enzymatic" and "non-enzymatic" types of system with or without the presence of oxygen or co-factors.Cnidaria: A phylum of radially symmetrical invertebrates characterized by possession of stinging cells called nematocysts. It includes the classes ANTHOZOA; CUBOZOA; HYDROZOA, and SCYPHOZOA. Members carry CNIDARIAN VENOMS.Thoracica: A superorder of marine CRUSTACEA, free swimming in the larval state, but permanently fixed as adults. There are some 800 described species, grouped in several genera, and comprising of two major orders of barnacles: stalked (Pedunculata) and sessile (Sessilia).Papillary Muscles: Conical muscular projections from the walls of the cardiac ventricles, attached to the cusps of the atrioventricular valves by the chordae tendineae.Carnivora: An order of MAMMALS, usually flesh eaters with appropriate dentition. Suborders include the terrestrial carnivores Fissipedia, and the aquatic carnivores PINNIPEDIA.Apoproteins: The protein components of a number of complexes, such as enzymes (APOENZYMES), ferritin (APOFERRITINS), or lipoproteins (APOLIPOPROTEINS).Bupranolol: An adrenergic-beta-2 antagonist that has been used for cardiac arrhythmia, angina pectoris, hypertension, glaucoma, and as an antithrombotic.Luminescent Agents: Compound such as LUMINESCENT PROTEINS that cause or emit light (PHYSICAL LUMINESCENCE).Caffeine: A methylxanthine naturally occurring in some beverages and also used as a pharmacological agent. Caffeine's most notable pharmacological effect is as a central nervous system stimulant, increasing alertness and producing agitation. It also relaxes SMOOTH MUSCLE, stimulates CARDIAC MUSCLE, stimulates DIURESIS, and appears to be useful in the treatment of some types of headache. Several cellular actions of caffeine have been observed, but it is not entirely clear how each contributes to its pharmacological profile. Among the most important are inhibition of cyclic nucleotide PHOSPHODIESTERASES, antagonism of ADENOSINE RECEPTORS, and modulation of intracellular calcium handling.Strophanthidin: 3 beta,5,14-Trihydroxy-19-oxo-5 beta-card-20(22)-enolide. The aglycone cardioactive agent isolated from Strophanthus Kombe, S. gratus and other species; it is a very toxic material formerly used as digitalis. Synonyms: Apocymarin; Corchorin; Cynotoxin; Corchorgenin.PyrazinesFirefly Luciferin: A benzothaizole which is oxidized by LUCIFERASES, FIREFLY to cause emission of light (LUMINESCENCE).Arsenazo III: Metallochrome indicator that changes color when complexed to the calcium ion under physiological conditions. It is used to measure local calcium ion concentrations in vivo.Egtazic Acid: A chelating agent relatively more specific for calcium and less toxic than EDETIC ACID.Myocardial Contraction: Contractile activity of the MYOCARDIUM.Ryanodine: A methylpyrrole-carboxylate from RYANIA that disrupts the RYANODINE RECEPTOR CALCIUM RELEASE CHANNEL to modify CALCIUM release from SARCOPLASMIC RETICULUM resulting in alteration of MUSCLE CONTRACTION. It was previously used in INSECTICIDES. It is used experimentally in conjunction with THAPSIGARGIN and other inhibitors of CALCIUM ATPASE uptake of calcium into SARCOPLASMIC RETICULUM.Mycorrhizae: Symbiotic combination (dual organism) of the MYCELIUM of FUNGI with the roots of plants (PLANT ROOTS). The roots of almost all higher plants exhibit this mutually beneficial relationship, whereby the fungus supplies water and mineral salts to the plant, and the plant supplies CARBOHYDRATES to the fungus. There are two major types of mycorrhizae: ectomycorrhizae and endomycorrhizae.Symbiosis: The relationship between two different species of organisms that are interdependent; each gains benefits from the other or a relationship between different species where both of the organisms in question benefit from the presence of the other.Fungi: A kingdom of eukaryotic, heterotrophic organisms that live parasitically as saprobes, including MUSHROOMS; YEASTS; smuts, molds, etc. They reproduce either sexually or asexually, and have life cycles that range from simple to complex. Filamentous fungi, commonly known as molds, refer to those that grow as multicellular colonies.Glomeromycota: A phylum of fungi that are mutualistic symbionts and form ARBUSCULAR MYCORRHIZAE with PLANT ROOTS.Cell-Penetrating Peptides: Peptides that have the ability to enter cells by crossing the plasma membrane directly, or through uptake by the endocytotic pathway.Daucus carota: A plant species of the family APIACEAE that is widely cultivated for the edible yellow-orange root. The plant has finely divided leaves and flat clusters of small white flowers.Plant Roots: The usually underground portions of a plant that serve as support, store food, and through which water and mineral nutrients enter the plant. (From American Heritage Dictionary, 1982; Concise Dictionary of Biology, 1990)Laboratories, Dental: Facilities for the performance of services related to dental treatment but not done directly in the patient's mouth.Dental Technicians: Individuals responsible for fabrication of dental appliances.Biology: One of the BIOLOGICAL SCIENCE DISCIPLINES concerned with the origin, structure, development, growth, function, genetics, and reproduction of animals, plants, and microorganisms.Technology, Dental: The field of dentistry involved in procedures for designing and constructing dental appliances. It includes also the application of any technology to the field of dentistry.Oral Medicine: A branch of dentistry dealing with diseases of the oral and paraoral structures and the oral management of systemic diseases. (Hall, What is Oral Medicine, Anyway? Clinical Update: National Naval Dental Center, March 1991, p7-8)Laboratories: Facilities equipped to carry out investigative procedures.Databases, Chemical: Databases devoted to knowledge about specific chemicals.Luciferases: Enzymes that oxidize certain LUMINESCENT AGENTS to emit light (PHYSICAL LUMINESCENCE). The luciferases from different organisms have evolved differently so have different structures and substrates.Adenosine Triphosphate: An adenine nucleotide containing three phosphate groups esterified to the sugar moiety. In addition to its crucial roles in metabolism adenosine triphosphate is a neurotransmitter.Therapies, Investigational: Treatments which are undergoing clinical trials or for which there is insufficient evidence to determine their effects on health outcomes; coverage for such treatments is often denied by health insurers.Cystic Fibrosis: An autosomal recessive genetic disease of the EXOCRINE GLANDS. It is caused by mutations in the gene encoding the CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR expressed in several organs including the LUNG, the PANCREAS, the BILIARY SYSTEM, and the SWEAT GLANDS. Cystic fibrosis is characterized by epithelial secretory dysfunction associated with ductal obstruction resulting in AIRWAY OBSTRUCTION; chronic RESPIRATORY INFECTIONS; PANCREATIC INSUFFICIENCY; maldigestion; salt depletion; and HEAT PROSTRATION.Bronchi: The larger air passages of the lungs arising from the terminal bifurcation of the TRACHEA. They include the largest two primary bronchi which branch out into secondary bronchi, and tertiary bronchi which extend into BRONCHIOLES and PULMONARY ALVEOLI.Type C Phospholipases: A subclass of phospholipases that hydrolyze the phosphoester bond found in the third position of GLYCEROPHOSPHOLIPIDS. Although the singular term phospholipase C specifically refers to an enzyme that catalyzes the hydrolysis of PHOSPHATIDYLCHOLINE (EC 3.1.4.3), it is commonly used in the literature to refer to broad variety of enzymes that specifically catalyze the hydrolysis of PHOSPHATIDYLINOSITOLS.Epithelial Cells: Cells that line the inner and outer surfaces of the body by forming cellular layers (EPITHELIUM) or masses. Epithelial cells lining the SKIN; the MOUTH; the NOSE; and the ANAL CANAL derive from ectoderm; those lining the RESPIRATORY SYSTEM and the DIGESTIVE SYSTEM derive from endoderm; others (CARDIOVASCULAR SYSTEM and LYMPHATIC SYSTEM) derive from mesoderm. Epithelial cells can be classified mainly by cell shape and function into squamous, glandular and transitional epithelial cells.Lung: Either of the pair of organs occupying the cavity of the thorax that effect the aeration of the blood.Anti-Inflammatory Agents, Non-Steroidal: Anti-inflammatory agents that are non-steroidal in nature. In addition to anti-inflammatory actions, they have analgesic, antipyretic, and platelet-inhibitory actions.They act by blocking the synthesis of prostaglandins by inhibiting cyclooxygenase, which converts arachidonic acid to cyclic endoperoxides, precursors of prostaglandins. Inhibition of prostaglandin synthesis accounts for their analgesic, antipyretic, and platelet-inhibitory actions; other mechanisms may contribute to their anti-inflammatory effects.Wolfram Syndrome: A hereditary condition characterized by multiple symptoms including those of DIABETES INSIPIDUS; DIABETES MELLITUS; OPTIC ATROPHY; and DEAFNESS. This syndrome is also known as DIDMOAD (first letter of each word) and is usually associated with VASOPRESSIN deficiency. It is caused by mutations in gene WFS1 encoding wolframin, a 100-kDa transmembrane protein.Inositol 1,4,5-Trisphosphate Receptors: Intracellular receptors that bind to INOSITOL 1,4,5-TRISPHOSPHATE and play an important role in its intracellular signaling. Inositol 1,4,5-trisphosphate receptors are calcium channels that release CALCIUM in response to increased levels of inositol 1,4,5-trisphosphate in the CYTOPLASM.Endoplasmic Reticulum: A system of cisternae in the CYTOPLASM of many cells. In places the endoplasmic reticulum is continuous with the plasma membrane (CELL MEMBRANE) or outer membrane of the nuclear envelope. If the outer surfaces of the endoplasmic reticulum membranes are coated with ribosomes, the endoplasmic reticulum is said to be rough-surfaced (ENDOPLASMIC RETICULUM, ROUGH); otherwise it is said to be smooth-surfaced (ENDOPLASMIC RETICULUM, SMOOTH). (King & Stansfield, A Dictionary of Genetics, 4th ed)Mitochondria: Semiautonomous, self-reproducing organelles that occur in the cytoplasm of all cells of most, but not all, eukaryotes. Each mitochondrion is surrounded by a double limiting membrane. The inner membrane is highly invaginated, and its projections are called cristae. Mitochondria are the sites of the reactions of oxidative phosphorylation, which result in the formation of ATP. They contain distinctive RIBOSOMES, transfer RNAs (RNA, TRANSFER); AMINO ACYL T RNA SYNTHETASES; and elongation and termination factors. Mitochondria depend upon genes within the nucleus of the cells in which they reside for many essential messenger RNAs (RNA, MESSENGER). Mitochondria are believed to have arisen from aerobic bacteria that established a symbiotic relationship with primitive protoeukaryotes. (King & Stansfield, A Dictionary of Genetics, 4th ed)Lipid Bilayers: Layers of lipid molecules which are two molecules thick. Bilayer systems are frequently studied as models of biological membranes.Chromosomes, Human, Pair 4: A specific pair of GROUP B CHROMOSOMES of the human chromosome classification.Zinostatin: An enediyne that alkylates DNA and RNA like MITOMYCIN does, so it is cytotoxic.

Intracellular trafficking pathways in the assembly of connexins into gap junctions. (1/417)

Trafficking pathways underlying the assembly of connexins into gap junctions were examined using living COS-7 cells expressing a range of connexin-aequorin (Cx-Aeq) chimeras. By measuring the chemiluminescence of the aequorin fusion partner, the translocation of oligomerized connexins from intracellular stores to the plasma membrane was shown to occur at different rates that depended on the connexin isoform. Treatment of COS-7 cells expressing Cx32-Aeq and Cx43-Aeq with brefeldin A inhibited the movement of these chimera to the plasma membrane by 84 +/- 4 and 88 +/- 4%, respectively. Nocodazole treatment of the cells expressing Cx32-Aeq and Cx43-Aeq produced 29 +/- 16 and 4 +/- 7% inhibition, respectively. In contrast, the transport of Cx26 to the plasma membrane, studied using a construct (Cx26/43T-Aeq) in which the short cytoplasmic carboxyl-terminal tail of Cx26 was replaced with the extended carboxyl terminus of Cx43, was inhibited 89 +/- 5% by nocodazole and was minimally affected by exposure of cells to brefeldin A (17 +/-11%). The transfer of Lucifer yellow across gap junctions between cells expressing wild-type Cx32, Cx43, and the corresponding Cx32-Aeq and Cx43-Aeq chimeras was reduced by nocodazole treatment and abolished by brefeldin A treatment. However, the extent of dye coupling between cells expressing wild-type Cx26 or the Cx26/43T-Aeq chimeras was not significantly affected by brefeldin A treatment, but after nocodazole treatment, transfer of dye to neighboring cells was greatly reduced. These contrasting effects of brefeldin A and nocodazole on the trafficking properties and intercellular dye transfer are interpreted to suggest that two pathways contribute to the routing of connexins to the gap junction.  (+info)

Reactive oxygen metabolites increase mitochondrial calcium in endothelial cells: implication of the Ca2+/Na+ exchanger. (2/417)

In endothelial cells, a bolus of hydrogen peroxide (H2O2) or oxygen metabolites generated by hypoxanthine-xanthine oxidase (HX-XO) increased the mitochondrial calcium concentration [Ca2+]m. Both agents caused a biphasic increase in [Ca2+]m which was preceded by a rise in cytosolic free calcium concentration [Ca2+]c (18 and 6 seconds for H2O2 and HX-XO, respectively). The peak and plateau elevations of [Ca2+] were consistently higher in the mitochondrial matrix than in the cytosol. In Ca2+-free/EGTA medium, the plateau phase of elevated [Ca2+] evoked by H2O2 due to capacitative Ca2+ influx was abolished in the cytosol, but was maintained in the mitochondria. In contrast to H2O2 and HX-XO, ATP which binds the P2Y purinoceptors induced an increase in [Ca2+]m that was similar to that of [Ca2+]c. When cells were first stimulated with inositol 1,4, 5-trisphosphate-generating agonists or the Ca2+-ATPase inhibitor cyclopiazonic acid (CPA), subsequent addition of H2O2 did not affect [Ca2+]c, but still caused an elevation of [Ca2+]m. Moreover, the specific inhibitor of the mitochondrial Ca2+/Na+ exchanger, 7-chloro-3,5-dihydro-5-phenyl-1H-4.1-benzothiazepine-2-on (CGP37157), did not potentiate the effects of H2O2 and HX-XO on [Ca2+]m, while causing a marked increase in the peak [Ca2+]m and a significant attenuation of the rate of [Ca2+]m efflux upon addition of histamine or CPA. In permeabilized cells, H2O2 mimicked the effects of CGP37157 causing an increase in the basal level of matrix free Ca2+ and decreased efflux. Dissipation of the electrochemical proton gradient by carbonylcyanide p-(trifluoromethoxy) phenylhydrazone (FCCP), and blocade of the Ca2+ uptake by ruthenium red prevented [Ca2+]m increases evoked by H2O2. These results demonstrate that the H2O2-induced elevation in [Ca2+]m results from a transfer of Ca2+ secondary to increased [Ca2+]c, and an inhibition of the Ca2+/Na+ electroneutral exchanger of the mitochondria.  (+info)

Functional effects of endothelin and regulation of endothelin receptors in isolated human nonfailing and failing myocardium. (3/417)

BACKGROUND: An activated endothelin (ET) system may be of pathophysiological relevance in human heart failure. We characterized the functional effects of ET-1, ET receptors, and ET-1 peptide concentration in left ventricular myocardium from 10 nonfailing hearts (NF) and 27 hearts in end-stage failure due to idiopathic dilative cardiomyopathy (DCM). METHODS AND RESULTS: Inotropic effects were characterized in isolated muscle strips (1 Hz; 37 degrees C). ET-1 0.0001 to 0.3 micromol/L significantly (P<0.05) increased twitch force by maximally 59+/-10% in NF and by 36+/-11% in DCM (P<0.05 versus NF). Preincubation with propranolol 1 micromol/L and prazosin 0.1 micromol/L did not affect the response to ET-1, but the mixed ET receptor antagonist bosentan and the ETA receptor antagonist BQ-123 shifted the concentration-response curves for ET-1 rightward. The ETB receptor agonist sarafotoxin S6c 0.001 to 0.3 micromol/L had no functional effects. The inotropic response to ET-1 was not associated with increased intracellular Ca2+ transients, as assessed in aequorin-loaded muscle strips. ET receptor density (Bmax; radioligand binding) was 62.5+/-12.5 fmol/mg protein in NF and 122. 4+/-24.3 fmol/mg protein in DCM (P<0.05 versus NF). The increase in Bmax in DCM resulted from an increase in ETA receptors without change in ETB receptors. ET-1 peptide concentration (radioimmunoassay) was higher in DCM than in NF (14 447+/-2232 versus 4541+/-1340 pg/mg protein, P<0.05). CONCLUSIONS: ET-1 exerts inotropic effects in human myocardium through ETA receptor-mediated increases in myofibrillar Ca2+ responsiveness. In DCM, functional effects of ET-1 are attenuated, but ETA receptor density and ET-1 peptide concentration are increased, indicating an activated local cardiac ET system and possibly a reduced postreceptor signaling efficiency.  (+info)

Secretagogues modulate the calcium concentration in the endoplasmic reticulum of insulin-secreting cells. Studies in aequorin-expressing intact and permeabilized ins-1 cells. (4/417)

The precise regulation of the Ca2+ concentration in the endoplasmic reticulum ([Ca2+]er) is important for protein processing and signal transduction. In the pancreatic beta-cell, dysregulation of [Ca2+]er may cause impaired insulin secretion. The Ca2+-sensitive photoprotein aequorin mutated to lower its Ca2+ affinity was stably expressed in the endoplasmic reticulum (ER) of rat insulinoma INS-1 cells. The steady state [Ca2+]er was 267 +/- 9 microM. Both the Ca2+-ATPase inhibitor cyclopiazonic acid and 4-chloro-m-cresol, an activator of ryanodine receptors, caused an almost complete emptying of ER Ca2+. The inositol 1,4,5-trisphosphate generating agonists, carbachol, and ATP, reduced [Ca2+]er by 20-25%. Insulin secretagogues that raise cytosolic [Ca2+] by membrane depolarization increased [Ca2+]er in the potency order K+ >> glucose > leucine, paralleling their actions in the cytosolic compartment. Glucose, which augmented [Ca2+]er by about 25%, potentiated the Ca2+-mobilizing effect of carbachol, explaining the corresponding observation in cytosolic [Ca2+]. The filling of ER Ca2+ by glucose is not directly mediated by ATP production as shown by the continuous monitoring of cytosolic ATP in luciferase expressing cells. Both glucose and K+ increase [Ca2+]er, but only the former generated whereas the latter consumed ATP. Nonetheless, drastic lowering of cellular ATP with a mitochondrial uncoupler resulted in a marked decrease in [Ca2+]er, emphasizing the requirement for mitochondrially derived ATP above a critical threshold concentration. Using alpha-toxin permeabilized cells in the presence of ATP, glucose 6-phosphate did not change [Ca2+]er, invalidating the hypothesis that glucose acts through this metabolite. Therefore, insulin secretagogues that primarily stimulate Ca2+ influx, elevate [Ca2+]er to ensure beta-cell homeostasis.  (+info)

Reactive oxygen species activate a Ca2+-dependent cell death pathway in the unicellular organism Trypanosoma brucei brucei. (5/417)

Here we examine a cell death process induced by reactive oxygen species (ROS) in the haemoflagellate Trypanosoma brucei brucei. Ca2+ distribution in cellular compartments was measured with stable transformants expressing aequorin targeted to the cytosol, nucleus or mitochondrion. Within 1.5 h of ROS production, mitochondrial Ca2+ transport was impaired and the Ca2+ barrier between the nuclear envelope and cytosol was disrupted. Consequently the mitochondrion did not accumulate Ca2+ efficiently in response to an extracellular stimulus, and excess Ca2+ accumulated in the nucleus. The terminal transferase deoxytidyl uridine end labelling assay revealed that, 5 h after treatment with ROS, extensive fragmentation of nuclear DNA occurred in over 90% of the cells. Permeability changes in the plasma membrane did not occur until an additional 2 h had elapsed. The intracellular Ca2+ buffer, EGTA acetoxymethyl ester, prevented DNA fragmentation and prolonged the onset of changes in cell permeability. Despite some similarities to apoptosis, nuclease activation was not a consequence of caspase 3, caspase 1, calpain, serine protease, cysteine protease or proteasome activity. Moreover, trypanosomes expressing mouse Bcl-2 were not protected from ROS even though protection from mitochondrial dysfunction and ROS have been reported for mammalian cells. Overall, these results demonstrate that Ca2+ pathways can induce pathology in trypanosomes, although the specific proteins involved might be distinct from those in metazoans.  (+info)

The vacuolar Ca2+/H+ exchanger Vcx1p/Hum1p tightly controls cytosolic Ca2+ levels in S. cerevisiae. (6/417)

It is well established that the vacuole plays an important role in the cellular adaptation to growth in the presence of elevated extracellular Ca2+ concentrations in Saccharomyces cerevisiae. The Ca2+ ATPase Pmc1p and the Ca2+/H+ exchanger Vcx1p/Hum1p have been shown to facilitate Ca2+ sequestration into the vacuole. However, the distinct physiological roles of these two vacuolar Ca2+ transporters remain uncertain. Here we show that Vcx1p can rapidly sequester a sudden pulse of cytosolic Ca2+ into the vacuole, while Pmc1p carries out this function much less efficiently. This finding is consistent with the postulated role of Vcx1p as a high capacity, low affinity Ca2+ transporter and suggests that Vcx1p may act to attenuate the propagation of Ca2+ signals in this organism.  (+info)

Differential pharmacological properties and signal transduction of the sphingosine 1-phosphate receptors EDG-1, EDG-3, and EDG-5. (7/417)

Sphingosine 1-phosphate (SPP) is a potent lipid mediator released upon cellular activation. In this report, pharmacological properties of the three G-protein-coupled receptors (GPCRs) for SPP, EDG-1, -3, and -5 are characterized using a Xenopus oocyte expression system, which lacks endogenous SPP receptors. Microinjection of the EDG-3 and EDG-5 but not EDG-1 mRNA conferred SPP-responsive intracellular calcium transients; however, the EDG-5 response was quantitatively much less. Co-expression of EDG-1 receptor with the chimeric Galphaqi protein conferred SPP responsiveness. Galphaqi or Galphaq co-injection also potentiated the EDG-5 and EDG-3 mediated responses to SPP. These data suggest that SPP receptors couple differentially to the Gq and Gi pathway. All three GPCRs were also activated by sphingosylphosphorylcholine, albeit at higher concentrations. None of the other related sphingolipids tested stimulated or blocked SPP-induced calcium responses. However, suramin, a polycyclic anionic compound, selectively antagonized SPP-activated calcium transients in EDG-3 expressing oocytes with an IC50 of 22 microM, suggesting that it is an antagonist selective for the EDG-3 GPCR isotype. We conclude that the three SPP receptors signal differentially by coupling to different G-proteins. Furthermore, because only EDG-3 was antagonized by suramin, variations in receptor structure may determine differences in antagonist selectivity. This property may be exploited to synthesize receptor subtype-specific antagonists.  (+info)

Measurement of perimitochondrial Ca2+ concentration in bovine adrenal glomerulosa cells with aequorin targeted to the outer mitochondrial membrane. (8/417)

Microdomains of high cytosolic free Ca(2+) concentration in the proximity of mitochondria might have an important role in the stimulation of steroidogenesis in bovine adrenal glomerulosa cells. In the present study we have investigated local changes of free Ca(2+) concentration near the outer mitochondrial membrane ([Ca(2+)](om)) under stimulation with angiotensin II (Ang II) and K(+). Glomerulosa cells in primary culture were transfected with a recombinant cDNA encoding the N-terminal region of the human translocase protein 20 of the outer mitochondrial membrane, in frame with the Ca(2+)-sensitive photoprotein aequorin. This chimaeric aequorin (TomAeq) was associated with mitochondria-enriched subcellular fractions of transfected COS-7 cells and was susceptible to proteinase K, showing that it was targeted to the outer mitochondrial membrane, facing the cytosolic space. In bovine adrenal glomerulosa cells transfected with TomAeq cDNA, Ang II induced a transient [Ca(2+)](om) peak reaching 1.42+/-0.28 microM, which decreased immediately to the basal resting value. The peak response to Ang II was strikingly lower than the peak response of mitochondrial free Ca(2+) concentration, which increased to 5.4+/-1.2 microM. The smaller response of [Ca(2+)](om) to Ang II compared with the elevated matrix response did not result from buffering effects of the organelle, from altered mechanisms of intramitochondrial Ca(2+) transport or from differences in the affinity of the chimaeric aequorins for Ca(2+). This approach has allowed us to follow perimitochondrial Ca(2+) homeostasis in bovine glomerulosa cells under stimulation with Ca(2+)-mobilizing agonists and to reveal a strong gradient of Ca(2+) concentration between the mitochondrial matrix and the immediate environment of the organelle.  (+info)

Aequorin, which luminesces in the presence of calcium, was injected into photoreceptor cells of Limulus ventral eye. A bright light stimulus elicited a large increase in aequorin luminescence, the aequorin response, indicating a rise of intracellular calcium ion concentration, Cai. The aequorin response reached a maximum after the peak of the electrical response of the photoreceptor, decayed during a prolonged stimulus, and returned to an undetectable level in the dark. Reduction of Cao reduced the amplitude of the aequorin response by a factor no greater than 3. Raising Cao increased the amplitude of the aequorin response. The aequorin response became smaller when membrane voltage was clamped to successively more positive values. These results indicate that the stimulus-induced rise of Cai may be due in part to a light-induced influx of Ca and in part to release of Ca from an intracellular store. Our findings are consistent with the hypothesis that a rise in Cai is a step in the sequence of ...
Light emitting molecules are an indispensable part of detection and reporting in many fields and are employed in a variety of biomedical applications. Bioluminescent light, or living light, from bioluminescent proteins in particular has many beneficial characteristics, including their lack of a need for an outside excitation source and detection at as low as subattomole levels. Aequorin is a well-characterized bioluminescent photoprotein that has found application in in vitro and in vivo studies. Despite the many advantages of aequorin, its application has been limited by the finite number of canonical amino acids restricting the engineering of aequorin. In order to increase the applications of aequorin, we have taken established methods that hijack the cellular machinery used to synthesize proteins to incorporate non-natural amino acids. By site-specifically incorporating the non-natural amino acids L-4-aminophenylalanine, L-4-bromophenylalanine, L-4-iodophenylalanine, and L-4-methoxyphenylalanine,
... INTRODUCTION ...Many G-protein coupled receptors (GPCRs) trigger upon binding of an a...One of the methods of choice (reviewed by Mottheakis and Ohler 2000) ... ...,Cell,lines,expressing,recombinant,aequorin,and,a,G-protein,coupled,receptor,for,functional,screening,biological,advanced biology technology,biology laboratory technology,biology device technology,latest biology technology
Growth patterns and intracellular Ca2+ concentrations in the mutant strain Aspergillus awamori 66A containing a recombinant aequorin gene were studied in the presence of a permeabilizing fungicidal agent amphotericin B. The cell response, i.e., changes in the growth and development of the fungus (initiation of spore germination, mycelial growth, and intensity of sporulation) was dose-dependent. Low concentrations of amphotericin B (2.5 μM) stimulated spore germination: the number of germinating spores was 2-3 times higher than in the control (without the fungicide). At higher amphotericin concentrations (20 μM) spore germination was inhibited. Amphotericin B had a dose-dependent effect on mycelial growth and sporulation intensity on solid Vogel medium. Intracellular Ca2+ concentrations in the presence of amphotericin B were investigated using the luminescence of the photoprotein aequorin. High concentrations of amphotericin B (10 and 20 μM) were shown to cause an instantaneous increase in Ca2+
Aequorin is a protein that contains coelenterazine as a luminescent compound and can be used for intracellular calcium ion detection. When three Ca2+ ions bind to the aequorin complex consisting of the 22 KD apoaequorin protein (APO), molecular oxygen and the luminophore coelenterazine, the latter is oxidized to coelenteramide with a concomitant release of CO2 and blue light. Since the intensity of bioluminescence emitted by aequorin upon calcium binding correlates with the Ca2+ concentration, a sensitive measurement of Ca2+ concentrations with a broad detection range (~0.1 µM to ,100 µM) is possible. Unlike fluorescent Ca2+ indicators, Ca2+-bound aequorin can be detected without illuminating the sample, thereby eliminating interference from autofluorescence ...
Ca2+ is a key regulator not only of multiple cytosolic enzymes, but also of a variety of metabolic pathways occurring within the lumen of intracellular organelles. Until recently, no technique to selectively monitor the Ca2+ concentration within defined cellular compartments was available. We have recently proposed the use of molecularly engineered Ca(2+)-sensitive photoproteins to obtain such a result and demonstrated the application of this methodology to the study of mitochondrial and nuclear Ca2+ dynamics. We here describe in more detail the use of chimeric recombinant aequorin targeted to the mitochondria. The technique can be applied with equivalent results to different cell models, transiently or permanently transfected. In all the cell types we analyzed, mitochondrial Ca2+ concentration ([Ca2+]m) increases rapidly and transiently upon stimulation with agonists coupled to InsP3 generation. We confirm that the high speed of mitochondrial Ca2+ accumulation with this type of stimuli depends ...
Quincy Biosciecne Products Quincy Bioscience researches, develops and plans to market therapeutic compounds for the treatment of neurodegenerative disorders, such as Alzheimer s disease, Parkinson s disease, stroke, Huntington s disease, and Amyotrophic Lateral Sclerosis (ALS). New therapeutics will be based on the calcium-binding protein aequorin. Calcium-binding proteins are important in the protection of certain cellular populations and decrease over the course of aging or disease progression. In June 2004, the company applied for its first patent on the aequorin technology. Research and development work is conducted in university laboratories in addition to Quincy Biosciences own facilities to fully develop the therapeutic applications of this technology. Aequorin acts in a protective manner similar to proteins that are depleted in humans with these diseases of aging. The dietary supplement Prevagen has been developed based on Quincy Bioscience s core technology and is slated for launch ...
Single rat hepatocytes microinjected with aequorin generate oscillations in cytosolic free Ca2+ concentration ([Ca2+]i) when stimulated with agonists acting through the phosphoinositide signalling pathway. The duration of these transients has been shown to be characteristic of the stimulating agonist, so that transients of very different duration can be induced in the same individual hepatocyte by different agonists. In a previous study we have shown that ADP and ATP, which are believed to act through a single P2y-purinoceptor species, elicit very different [Ca2+]i responses in most of the hepatocytes. We have interpreted this as evidence for two Ca(2+)-mobilizing purinoceptors. The methylated derivative of ATP, adenosine 5′-[alpha beta-methylene]-triphosphate (pp[CH2]pA), is only a weak P2y-purinoceptor agonist. When 100 microM pp[CH2]pA was supplied to aequorin-injected hepatocytes, there was no effect on [Ca2+]i. However, 25 microM pp[CH2]pA co-supplied with ATP causes a potentiation of the ...
Rapid transient elevation of cytoplasmic calcium (Ca2+) levels in plant cells is an early signaling event triggered by many environmental cues including abiotic and biotic stresses. Cellular Ca2+ levels and their alterations can be monitored by genetically encoded reporter systems such as the bioluminescent protein, aequorin. Employment of proteinaceous Ca2+ sensors is usually performed in transgenic lines that constitutively express the reporter construct. Such settings limit the usage of these Ca2+ biosensors to particular reporter variants and plant genetic backgrounds, which can be a severe constraint in genetic pathway analysis. Here we systematically explored the potential of Arabidopsis thaliana leaf mesophyll protoplasts, either derived from a transgenic apoaequorin-expressing line or transfected with apoaequorin reporter constructs, as a complementary biological resource to monitor cytoplasmic changes of Ca2+ levels in response to various biotic stress elicitors. We tested a range of ...
The subplasmalemmal spatiotemporal aspects of Ca2+ dynamics sensitive to [Ca2+]o were examined using cameleon and a FRET-based technique. Our observations appear to be consistent with those of a previous study using aequorin, in which a dramatic increase in [Ca2+]spm in A7r5 cells was detectable only when the sensor was expressed on the cytoplasmic face of the plasma membrane.3 Cameleon is superior to aequorin in terms of its imaging feasibility based on the intense fluorescence emitted from GFP mutants; consequently, subplasmalemmal Ca2+ waves were observed and characterized for the first time in this study. These subcortical Ca2+ waves cannot be detected by conventional imaging methods using indicators such as Indo-1, Fluo-4, or native YC that are nonspecifically loaded or expressed in the cytosol. Bulk cytosolic Ca2+ waves, which have been well described, basically consist of Ca2+ released from internal stores. Unlike the bulk cytosolic Ca2+ waves, the Ca2+ source for subplasmalemmal waves is ...
The subplasmalemmal spatiotemporal aspects of Ca2+ dynamics sensitive to [Ca2+]o were examined using cameleon and a FRET-based technique. Our observations appear to be consistent with those of a previous study using aequorin, in which a dramatic increase in [Ca2+]spm in A7r5 cells was detectable only when the sensor was expressed on the cytoplasmic face of the plasma membrane.3 Cameleon is superior to aequorin in terms of its imaging feasibility based on the intense fluorescence emitted from GFP mutants; consequently, subplasmalemmal Ca2+ waves were observed and characterized for the first time in this study. These subcortical Ca2+ waves cannot be detected by conventional imaging methods using indicators such as Indo-1, Fluo-4, or native YC that are nonspecifically loaded or expressed in the cytosol. Bulk cytosolic Ca2+ waves, which have been well described, basically consist of Ca2+ released from internal stores. Unlike the bulk cytosolic Ca2+ waves, the Ca2+ source for subplasmalemmal waves is ...
How researchers are applying new methods for protein tagging to monitor GPCR internalization and to search for drugs that interfere with receptor trafficking. Bioluminescent protein tagging methods simplify and expedite screening for drugs that modulate hunger and energy homeostasis.
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The protein encoded by this gene is a G-protein coupled receptor involved in the regulation of feeding behavior. The encoded protein binds the hypothalamic neuropeptides
Bioluminescent proteins (BLPs) widely exist in many living organisms. As BLPs are featured by the capability of emitting lights, they can be served as biomarkers and easily detected in biomedical research, such as gene expression analysis and signal transduction pathways. Therefore, accurate identification of BLPs is important for disease diagnosis and biomedical engineering. In this paper, we propose a novel accurate sequence-based method named PredBLP (Prediction of BioLuminescent Proteins) to predict BLPs. We collect a series of sequence-derived features, which have been proved to be involved in the structure and function of BLPs. These features include amino acid composition, dipeptide composition, sequence motifs and physicochemical properties. We further prove that the combination of four types of features outperforms any other combinations or individual features. To remove potential irrelevant or redundant features, we also introduce Fisher Markov Selector together with Sequential Backward
Green Fluoroscent Protein:. · The GFP was first discovered from the jellyfish Aequorea victoria. But it is also present in other organisms.. · The Aequorea victoria contains two luminescent proteins.. 1) Aequorin. 2) GFP, i.e. Green Fluoroscent Protein. · Aequorin, when interacts with Ca+2, it emits flashes of the blue light.. · The GFP aquires energy from Aequorin and emits green light.. Structure of GFP:. · The GFP from Aequorea victoria has an 11 stranded beta-barrel structure, with a alpha-helix running up the axis of the barrel.. · The chromophore is attached to alpha-helix in the center of barrel, few amino acids make the chromophore.. · The chromophore has Serine, Tyrosine and Glycine at the position 65; 66 and 67 respectively.. · 4-(p-hydroxybenzylidene)-imidazolidin-5-one attached to protein backbone through 1 and 2 position of the ring.. · Chromophore has hydrogen-bonding with amino acid residue and water molecules.. ...
BioAssay record AID 632132 submitted by ChEMBL: Antagonist activity at rat MCHR1 expressed in HEK293 cells by aequorin bioluminescence assay.
Safety assessment of Apoaequorin, a protein preparation: subchronic toxicity study in rats. - Daniel L Moran, Palma Ann Marone, Mark R Bauter, Madhu G Soni
Direct knowledge of Ca2+ patterns in vertebrate development is largely restricted to early stages, in which they control fertilization, ooplasmic segregation and cleavage. To explore new roles of Ca2+ in vertebrate development, we injected the Ca2+ indicator aequorin into zebrafish eggs and imaged Ca2+ throughout the first day of development. During early cleavages, a high Ca2+ zone is seen in the cleavage furrows. The high Ca2+ zone during first cleavage spreads as a slow wave (0.5 microm/second) and is preceded by three Ca2+ pulses within the animal pole region of the egg. When Ca2+ concentrations are clamped at the resting level by BAPTA buffer injection into the zygote, all signs of development are blocked. In later development, Ca2+ patterns are associated with cell movements during gastrulation, with neural induction, with brain regionalization, with formation of the somites and neural keel, with otic placode formation, with muscle movements and with formation of the heart. Particularly ...
Prevagen has been touted as a dietary supplement that helps to improve memory. Does Prevagen work?. Prevagen is a dietary supplement made by Quincy Bioscience, a company based in Wisconsin. It contains apoaequorin, a protein derived from a jellyfish. Evidence cited by Quincy Bioscience that Prevagen is effective is reported in a study in 2016. This study was not published in any journal.. The goal of the study was to determine whether the active ingredient of Prevagen, apoaequorin, improves cognitive function in older adults who have normal or near normal memory function. In this study, healthy adults ages 40-91 with memory concerns were examined by cognitive tests over a period of 90 days. People with neurological diseases and other conditions were excluded from the study population.. The authors reported that there were significant improvements in cognitive function in people treated with the study chemical compared to those who were not.. Limitations of this study include:. The study duration ...
Prevagen has been touted as a dietary supplement that helps to improve memory. Does Prevagen work?. Prevagen is a dietary supplement made by Quincy Bioscience, a company based in Wisconsin. It contains apoaequorin, a protein derived from a jellyfish. Evidence cited by Quincy Bioscience that Prevagen is effective is reported in a study in 2016. This study was not published in any journal.. The goal of the study was to determine whether the active ingredient of Prevagen, apoaequorin, improves cognitive function in older adults who have normal or near normal memory function. In this study, healthy adults ages 40-91 with memory concerns were examined by cognitive tests over a period of 90 days. People with neurological diseases and other conditions were excluded from the study population.. The authors reported that there were significant improvements in cognitive function in people treated with the study chemical compared to those who were not.. Limitations of this study include:. The study duration ...
FDSS Application Note No.1 Chloride ion sensitive fluorescent dye for Drug Screening [0.1MB/PDF]. FDSS Application Note No.2 CoroNa Red as a potential probe for intracellular sodium imaging [0.1MB/PDF]. FDSS Application Note No.3 FRET-based Voltage Sensor dyes for Drug Screening [0.1MB/PDF]. FDSS Application Note No.4 FDSS Cell dispensing unit for Aequorin Flash luminescence [0.4MB/PDF]. FDSS Application Note No.5 COX flash luminescence screening [0.1MB/PDF]. FDSS Application Note No.6 Direct comparison of Fast response and Slow response Membrane potential dye using the FDSS6000 [0.1MB/PDF]. FDSS Application Note No.7 Multiplexing Calcium Mobilization and Membrane Potential Assays Using the FDSS6000 [0.3MB/PDF]. FDSS Application Note No.8 Comparison of No Wash Reagent Kits on the FDSS6000 [0.4MB/PDF]. FDSS Application Note No.9 Effect of Plate Mixing, Fluid Addition Height and Speed on Reducing Addition Artifacts and Negative Control Drift Using the FDSS6000 [0.1MB/PDF]. FDSS Application Note ...
Mark Underwood of Quincy Bioscience figured out that giving aequorin to rats through the golden months of their lives helped them hang onto the ability to perform tricks despite their advancing age. Rats normally get less tricky as they age because they lose their calcium-binding proteins, allowing free calcium to ravage their brains, which lets the tricks leak out. But rats plus jellyfish protein equals peak performance at mazes and bells throughout the lifespan ...
The role of Na(+)-Ca2+ exchange in paired pulse potentiation of ferret ventricular muscle.: 1. Stimulation of cardiac muscle with pairs of stimuli (paired puls
The appeal that Prevagen is making to consumers is a very old one: basically, they want you to think that if a protein is used in your brain, then eating that protein will make your brain healthier (even though apoaequorin is not a human protein). By this argument, I could package up hundreds of different proteins (perhaps thousands-the brain is a complex organ) and sell them as "brain food" This simplistic principle has been used for centuries in folk medicine: its the reason why some people think that eating the body parts of bears and tigers will make them more virile. But eating tiger organs doesnt make you more like a tiger. Its a form of magical thinking, and theres simply no science to support it. In short, its just wrong ...
The appeal that Prevagen is making to consumers is a very old one: basically, they want you to think that if a protein is used in your brain, then eating that protein will make your brain healthier (even though apoaequorin is not a human protein). By this argument, I could package up hundreds of different proteins (perhaps thousands-the brain is a complex organ) and sell them as "brain food" This simplistic principle has been used for centuries in folk medicine: its the reason why some people think that eating the body parts of bears and tigers will make them more virile. But eating tiger organs doesnt make you more like a tiger. Its a form of magical thinking, and theres simply no science to support it. In short, its just wrong ...
Life, on the most basic level, is incredibly simple in theory yet vastly complex in practice. DNA is made out of only four nucleic acids: adenine which pai
The ability of labeled DNA or RNA probes to bind with high affinity and specificity to complementary nucleic acid sequences forms the basis of nucleic acid hybridization assays. Immunoassays, on the other hand, exploit the strong and specific interaction between antigens and antibodies. One aspect of this dissertation deals with enhancement of the sensitivity of bioluminescence hybridization assays based on the photoprotein aequorin. This is achieved by introducing, enzymatically, multiple aequorin labels per DNA hybrid. The bound aequorin is determined by its characteristic bioluminescence. An 8-fold improvement in sensitivity is observed with the amplified assay as compared to an assay without the amplification step. Another aspect of this dissertation involves the development of two novel microtiter well-based DNA hybridization assays in which an expressible cDNA fragment encoding firefly luciferase serves as a reporter molecule. The reporter molecule undergoes in vitro expression through coupled
Comments, concepts and statistics about Genomic organization, evolution, and expression of photoprotein and opsin genes in Mnemiopsis leidyi: a new view of ctenophore photocytes.
Exposure of the yeast Saccharomyces cerevisiae to alkaline stress resulted in adaptive changes that involved remodeling the gene expression. Recent evidence suggested that the calcium-activated protein phosphatase calcineurin could play a role in alkaline stress signaling. By using an aequorin luminescence reporter, we showed that alkaline stress resulted in a sharp and transient rise in cytoplasmic calcium. This increase was largely abolished by addition of EGTA to the medium or in cells lacking Mid1 or Cch1, components of the high affinity cell membrane calcium channel. Under these circumstances, the alkaline response of different calcineurin-sensitive transcriptional promoters was also blocked. Therefore, exposure to alkali resulted in entry of calcium from the external medium, and this triggered a calcineurin-mediated response. The involvement of calcineurin and Crz1/Tcn1, the transcription factor activated by the phosphatase, in the transcriptional response triggered by alkalinization has ...
Bioluminescence resonance energy transfer (BRET) is a well-established method for investigating protein-protein interactions. Here we present a BRET approach to monitor ligand binding to G protein-coupled receptors (GPCRs) on the surface of living cells made possible by the use of fluorescent ligands in combination with a bioluminescent protein (NanoLuc) that can be readily expressed on the N terminus of GPCRs.. ...
Repolarization alternans has been considered a strong marker of electrical instability. The objective of this study was to investigate the hypothesis that ischemia-induced contrasting effects on the kinetics of membrane voltage and intracellular calcium transient (Ca(i)T) can explain the vulnerability of the ischemic heart to repolarization alternans. Ischemia-induced changes in action potential (AP) and Ca(i)T resulting in alternans were investigated in perfused Langendorff guinea pig hearts subjected to 10-15 min of global no-flow ischemia followed by 10-15 min of reperfusion. The heart was stained with 100 microl of rhod-2 AM and 25 microl of RH-237, and AP and Ca(i)T were simultaneously recorded with an optical mapping system of two 16 x 16 photodiode arrays. Ischemia was associated with shortening of AP duration (D) but delayed upstroke, broadening of peak, and slowed decay of Ca(i)T resulting in a significant increase of Ca(i)T-D. The changes in APD were spatially heterogeneous in contrast to a
MeSH-minor] Animals. Drug Administration Schedule. Enzyme Activation. Eukaryotic Initiation Factor-2 / antagonists & inhibitors. Female. Genes, Reporter. Genes, p53. Green Fluorescent Proteins. Humans. Injections, Spinal. Luminescent Proteins / analysis. Luminescent Proteins / genetics. Mice. Mice, Nude. Neoplasm Proteins / physiology. Proto-Oncogene Proteins p21(ras) / physiology. Signal Transduction. Transcription, Genetic. Tumor Cells, Cultured. Virus Replication. Xenograft Model Antitumor Assays. eIF-2 Kinase / antagonists & inhibitors. eIF-2 Kinase / ...
The invention relates to the staining of poly(amino acids), including peptides, polypeptides and proteins in gels and on solid supports, using neutral or anionic complexes of transition metals.
Thyroid Hormones , Tumor Markers , Reproductive Endocrinology , Diabetes , Infectious Disease , Cardiac Markers , Adrenal/Pituitary , Bone Metabolism , TDM , Anemia , LIA Other Analytes , Nucleic acid quantification , Aequorin Ca++ assays , Ion channels / GPCR´s , Two-Hybrid / Protein - protein interaction , Apoptosis / Cell viability / Cell cytotoxicity , ATP determination , Reactive oxygen species (ROS) ...
AequoScreen Double Transfected Cell Line: Human recombinant 5-HT3A receptor in aequorin HEK-293 host cell.. We provide two vials of cryopreserved cells (approximately 2.5 x 106 cells/vial), detailed product information including cell line properties, culture conditions and the pharmacological properties of the recombinant channel in a functional assay.. All cell lines are tested for the absence of mycoplasma.. Terms and conditions apply. Some of our receptors may be restricted for sale in specified countries. Please inquire at your local sales office for more information.. Features:. ...
These cells express a novel variant of clytin, a calcium-activated photoprotein, to enable sensitive luminescent detection of ligand-induced calcium flux. The clytin contains a mutation that increases its affinity for calcium to a level that permits detection of cytosolic calcium in many cells with greater sensitivity than other mitochondrially expressed photoproteins. Luminescent calcium assays offer several advantages over fluorescent calcium assays including increased sensitivity and lack of interference
AequoScreen parental cells stably expressing the mitochondrially targeted apoaequorin and the G-protein Ga16 for flash-luminescence assay. Recombinant, in CHO-K1 host cell. GPCRs of your choice can be transiently / stably transfected into this cell line. Following GPCR stimulation, increases in intracellular calcium enable measurement of the resulting flash luminescence signal. Two vials of cryopreserved cells are shipped per order. Terms and conditions apply. Please inquire at your local sales office for more information.. ...
Individuals think, well, its natural, so it needs to work. This substance from one includes an essence from jellyfish and appears to have some kind of enchanting, science, fantasy kind of idea that it has to aid. I find that entertaining since why would certainly you assume that a protein from a primitive life form like a jellyfish would certainly have anything to do with the human mind? The substance concerned, apoaequorin (noticable a-po-ah-kwor-in), is a protein as well as it is undoubtedly broken down in the belly as well as not also taken in as an intact particle. Its broken down into amino acids, so it actually can not have any result in the human.. ...
V. Viviani et al. [Biochemistry 38 (1999) 8271] were the first to succeed in cloning the red-emitting enzyme from the South American railroad worm, which is the only bioluminescent organism known to emit a red-colored light. The application of red bioluminescence has been our goal because the transmittance of longer-wavelength light is superior to that of the other colors for visualization of biological functions in living cells. Now, different color luciferases, which emit with wavelength maxima ranging from 400 to 630 nm, are available and are being used. For example, based on different color luciferases, Nakajima et al. developed a tricolor reporter in vitro assay system based on these different color luciferases in which the expression of three genes can be monitored simultaneously. On the other hand, bioluminescence resonance energy transfer (BRET) is a natural phenomenon caused by the intermolecular interaction between a bioluminescent protein and a fluorophore on a second protein, ...
Accepted worldwide for its reliability, the FDSS is capable of 1536-well format and high-sensitivity luminescence measurements. The FDSS (Functional Drug Screening System) series are designed for cell-based assays in the drug discovery field. These instruments optically detect intracellular reactions and biological signal transmissions, and are used as screening systems to discover new lead compounds which could be candidates for new drugs. The FDSS7000EX is our high-end model capable of handling 1536-well assays and measuring both fluorescence and luminescence, and is equipped with a variety of functions such as multiple washing. Many kinds of suspended/adherent cells real-time kinetic reactions can be measured and analyzed. Various optional parts such as FRET, robot connections, etc. are available. In addition, the FDSS7000EX is expandable for future upgrades. Applications Intracellular calcium ion Membrane potential Ion channel Aequorin Luciferase FRET Suspended cell Applications Do more ...
AequoScreen® Double Transfected Cell Line: Human recombinant P2Y11 receptor in aequorin 1321N1 host cell. We provide two vials of cryopreserved cells (approximately 2.5 x 106 cells/vial), detailed product information including cell line properties, culture conditions and the pharmacological properties of the recombinant receptor in a functional assay. All cell lines are tested for the absence of mycoplasma. Terms and conditions apply. Some of our receptors may be restricted for sale in specified countries. Please inquire at your local sales office for more information.. Features:. ...
This book is an authoritative monograph on the recent progresses in the chemistry of bioluminescence. This book provides a comprehensive overview on the past and the latest development in understanding the biochemical mechanisms on some 35 different types of luminous organisms, together with information helpful to students and researchers in an Appendix. It is the first and only book that provides chemical information on all currently known bioluminescence systems. Dr Shimomura is the leading practitioner in the field for the past half century, and is best known for his discovery of the jellyfish photoprotein aequorin and the green fluorescent protein. This book is the bible of bioluminescence, and is "a must read," not only for the students who study bioluminescence but also for those who work in various aspects relating to bioluminescence. This book will be an important source of chemical knowledge on bioluminescence for a long period of time in future. Fully revised since its publication in ...
Ref. S69-ARF) Circa £24,000 per annum Fixed-term, three-year appointment We have an exciting Postdoctoral Research position which will investigate the role of cytoplasmic and chloroplastic Ca2+ signalling in the timing and induction of flowering in Arabidopsis thaliana. This ambitious project will apply new, GM technologies for manipulating and imaging subcellular [Ca2+] in vivo, using both aequorin (photon-counting imaging) and cameleon-GFPs (laser scanning confocal microscopy). The molecular effects of altered Ca2+ signalling will be investigated using a variety of techniques, notably real time quantitative PCR. The successful applicant will integrate a new research group led by Dr. John Love, in the Plant & Microbial Sciences Division and the Bio-imaging suite of Exeter University. More details at http://biojobs.blogspot.com/2007/04/postdoctoral-research-position-school.html More ARABIDOPSIS jobs available at http://biojobs.blogspot.com/search/label/Arabidopsis%20System ...
AequoScreen® Double Transfected Cell Line: Rat Mas-related MrgB3 receptor in aequorin CHO-K1 host cell. We provide two vials of cryopreserved cells (approximately 2.5x106 cells/vial), detailed product information including cell line properties, culture conditions and the pharmacological properties of the recombinant channel in a functional assay. All cell lines are tested for the absence of mycoplasma. Terms and conditions apply. Some of our receptors may be restricted for sale in specified countries. Please inquire at your local sales office for more information.. Features:. ...
Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dyes functional group. See the product information sheets for specific reactive dyes more information.. Coelenterazines and D-luciferin. Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also ...
Amines, aminooxy (also known as oxylamine), hydrazide, azide, alkyne, BCN, and tyramide reactive dyes, as well as dye free acids, are generally stable in aqueous solution when stored at -20°C for 6-12 months or longer, as long as no compounds are present that may react with the dyes functional group. See the product information sheets for specific reactive dyes more information.. Coelenterazines and D-luciferin. Coelenterazines are stable in solid form when stored as recommended; they are not stable in aqueous solution. Concentrated coelenterazine stock solutions (typically 1-100 mg/mL) should be prepared in ethanol or methanol; do not use DMSO or DMF to dissolve coelenterazines, because these solvents will oxidize the compounds. Ethanol or methanol stocks of coelenterazine can be stored at -20°C or below for six months or longer; alcohol stocks may evaporate during storage, so use tightly sealing screw cap vials and wrap the vials with Parafilm for long term storage. Propylene glycol also ...
A light generating dry disposable device for determining the presence of analytes in a test sample is disclosed. The device comprises a first zone containing conjugated ligand which is capable of reacting with analytes in the test sample. The ligand is conjugated with a photoprotein or related enzyme. The device further comprises a second trapping zone comprising immobilized analyte. The device also includes a third zone containing a reporter system that activates light generation by the conjugate. The conjugates are maintained in the first zone such that they are removable from the first zone when reacted with the soluble analytes from the test sample passing through the first zone, but not removed from the second trapping zone in the absence of such analytes. The third zone contains material capable of reacting with the photoprotein- or enzyme-linked ligand to produce a light-emitting reaction which indicates the presence of the analyte being tested. The present invention provides dry flow through
Green fluorescent protein (GFP), molecular model. The molecule has a cylindrical structure formed from beta sheets (ribbons). GFP is found in the Pacific jellyfish Aequorea victoria. It fluoresces green when blue light is shone on it. GFP is widely used as a research tool in biology and medicine. The gene coding for it can be tagged to the genes of other proteins or viruses to study their movements within cells. They can also be used to tag cancer cells to track their spread through the body. - Stock Image F006/9343
What is the meaning and origin of this expression? I cannot find it on the net. :-| You caused more trouble than a bag full of ferrets... He may be madder than a bag full of ferrets...
Furthermore, some of the blue light released by aequorin in contact with calcium ions is absorbed by a green fluorescent ... Other cofactors may be required for the reaction, such as calcium (Ca2+) for the photoprotein aequorin, or magnesium (Mg2+) ... ISBN 978-1-139-45181-9. Shimomura, O. (August 1995). "A short story of aequorin". The Biological Bulletin. 189 (1): 1-5. doi: ... Shimomura, O.; Johnson, F.H.; Saiga, Y. (1962). "Extraction, purification and properties of aequorin, a bioluminescent protein ...
The blue light produced is in turn transduced to green by the now famous green fluorescent protein (GFP). Both aequorin and GFP ... The species is best known as the source of two proteins involved in bioluminescence, aequorin, a photoprotein, and green ... PDB: 1EMA​ Shimomura O (August 1995). "A short story of aequorin". The Biological Bulletin. 189 (1): 1-5. doi:10.2307/1542194. ... In 1967, Ridgeway and Ashley microinjected aequorin into single muscle fibers of barnacles, and observed transient calcium ion- ...
For example, the photoprotein aequorin produces a flash of light when luciferin and calcium are added, rather than the ... Because of the kinetically slow step, each aequorin molecule must "recharge" with another molecule of luciferin before it can ... Shimomura O, Johnson FH (1975). "Regeneration of the photoprotein aequorin". Nature. 256 (5514): 236-238. doi:10.1038/256236a0 ... often until the addition of another required factor such as Ca2+ in the case of aequorin. Shimomura, O. "Bioluminescence: ...
He is known for his work to clone and sequence the genes for the photoprotein aequorin and green fluorescent protein (GFP) and ... Prasher, D., McCann, R.O., Cormier, M.J., Cloning and expression of the cDNA coding for aequorin, a bioluminescent calcium- ... aequorin. Photochem. Photobiol., 49(4), 509-512 (1989). Prasher, D.C., O'Kane, D., Lee, J., Woodward, B., The lumazine protein ... where he identified the gene sequence for aequorin. He then joined the Biology Department of the Woods Hole Oceanographic ...
Aequorin has been incorporated into human B cell lines for the detection of pathogenic bacteria and viruses in what is referred ... GFP, like aequorin, produces a blue fluorescent signal, but without the required addition of an exogenous substrate. All that ... Aequorin is a photoprotein isolated from the bioluminescent jellyfish Aequorea victoria. Upon addition of calcium ions (Ca2+) ... In some instances, the signal only occurs when a secondary substrate is added to the bioassay (luxAB, Luc, and aequorin). For ...
In the 1960s and 1970s, GFP, along with the separate luminescent protein aequorin (an enzyme that catalyzes the breakdown of ... The purpose of both the (primary) bioluminescence (from aequorin's action on luciferin) and the (secondary) fluorescence of GFP ... GFP is co-expressed with aequorin in small granules around the rim of the jellyfish bell. The secondary excitation peak (480 nm ... Prendergast FG, Mann KG (Aug 1978). "Chemical and physical properties of aequorin and the green fluorescent protein isolated ...
In 1961, Osamu Shimomura extracted green fluorescent protein (GFP) and another bioluminescent protein, called aequorin, from ... Shimomura, O.; Johnson, F. H.; Saiga, Y. (1962). "Extraction, purification and properties of aequorin, a bioluminescent protein ...
Hastings, J.W.; Mitchell, G.W.; Mattingly, P.H.; Blinks, J.R.; Van Leeuwen, M. (1969). "Response of aequorin bioluminescence to ... aequorin), which alone emits blue light, to a secondary green emitter which they termed green fluorescent protein (GFP). Once ...
"Connexin-aequorin chimerae report cytoplasmic calcium environments along trafficking pathways leading to gap junction ... "Assembly of chimeric connexin-aequorin proteins into functional gap junction channels. Reporting intracellular and plasma ...
Alternatively, Fura-2 , Furaptra , Indo-1 and aequorin may be used. An acetomethoxy group obscures the part of the molecule ...
Aequorin is also a useful tool to indicate calcium level inside cells; however, it has some limitations, primarily is that its ...
Prendergast, FG; Mann, KG (1978). "Chemical and physical properties of aequorin and the green fluorescent protein isolated from ...
At the final step of these reactions, Ca2+ ions are released, and in the presence of aequorin, photons are emitted. Aequorin is ... Aequorin," Photochem. Photobiol., vol. 49, no. 4, 1989, pp. 509-512. Petrovick, Martha S., James D. Harper, Frances E. Nargi, ...
Aequorin-expressing yeast emits light under electric control.J Biotechnol. 2011 Mar 20;152(3):93-5. Official website J ... It is based on yeast cells expressing aequorin protein sensitive to change in intracellular calcium. Upon electrical ...
It is the prosthetic group in the protein aequorin responsible for the blue light emission. Dinoflagellate luciferin is a ...
Prendergast, Franklyn G.; Mann, Kenneth G. (1978-08-22). "Chemical and physical properties of aequorin and the green ...
Another protein, aequorin, found in certain jellyfish, produces blue light in the presence of calcium. It can be used in ...
... can be visualised with fluorescence microscopy by using aequorin as a reporter protein. The ...
"The effects of digitalis on intracellular calcium transients in mammalian working myocardium as detected with aequorin". ...
Knight, Marc R.; Campbell, Anthony K.; Smith, Steven M.; Trewavas, Anthony J. (1991-08-08). "Transgenic plant aequorin reports ... aequorin, to report calcium signalling in plants. Together they obtained funding, created the plants and showed that they could ...
Knight, M. R.; Campbell, A. K.; Smith, S. M.; Trewavas, A. J. (1991). "Transgenic plant aequorin reports the effects of touch ...
In addition, aequorin has been used for years as an indicator of Ca2+ and has been shown to be safe and well tolerated by cells ... Aequorin belongs to the EF-hand family of CaBPs, with EF-hand loops that are closely related to CaBPs in mammals. ... Aequorin is made up of two components - the calcium binding component apoaequorin (AQ) and the chemiluminescent molecule ... EPS15 homology (EH) domain - InterPro: IPR000261 Aequorin is a calcium binding protein (CaBP) isolated from the coelenterate ...
"Determination of affinity and activity of ligands at the human neuropeptide Y Y4 receptor by flow cytometry and aequorin ...
"Determination of affinity and activity of ligands at the human neuropeptide Y Y4 receptor by flow cytometry and aequorin ...
Toisc go bhfuil an liosta bunaithe ar an leagan Béarla i 'Wikipedia en' beidh an leagan Béarla ar dtús agus an leagan Gaeilge ina dhiaidh. Tá súil agam, nuair atá méid áirithe den liosta aistrithe go Gaeilge gur féidir liosta i nGaeilge a chumadh nó i nGaeilge agus i mBéarla. ...
Chemical characterization of aequorin indicates the protein is somewhat resilient to harsh treatments. Aequorin is heat ... Shimomura O, Inouye S, Musicki B, Kishi Y (1990). "Recombinant aequorin and recombinant semi-synthetic aequorins. Cellular Ca2+ ... making aequorin suitable as a (Ca2+ reporter) in plants, fungi, and mammalian cells. Aequorin has a number of advantages over ... not aequorin, although both originally derived from the same animal. Work on aequorin began with E. Newton Harvey in 1921. ...
Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium.. Knight MR1, ... Microinjected aequorin has been widely used for intracellular calcium measurement in animal cells, but its use in plants has ... Reconstituted aequorin is cytoplasmic and nonperturbing; measurements can be made on whole plants and a calcium indicator can ... Aequorin is a calcium-sensitive luminescent protein from the coelenterate Aequorea victoria (A. forskalea) which is formed from ...
The intracellular delivery of TAT-aequorin reveals calcium-mediated sensing of environmental and symbiotic signals by the ... The intracellular delivery of TAT-aequorin reveals calcium-mediated sensing of environmental and symbiotic signals by the ...
... aequorin,and,a,G-protein,coupled,receptor,for,functional,screening,biological,advanced biology technology,biology laboratory ... For t he detection of antagonists, CHO-5HT2B- Gα16-aequorin cells were incubated with various concentrations of the compounds ... After reconstitution of active aequorin, the cells are diluted 10 times before use, and 50 μl of the dilution (i.e. 25 000 ... Button, D. and Brownstein, M. (1993) Aequorin-expressing mammalian cell lines used to report Ca2+ mobilization. Cell Calcium 14 ...
Antagonist activity at rat MCHR1 expressed in HEK293 cells by aequorin bioluminescence assay. ...
Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus ... Aequorin is a Ca2+ sensitive photoprotein suitable to measure intracellular Ca2+ transients in mammalian cells. ... pH also affects aequorin luminescence at values below 7. For these reasons, experiments with aequorin need to be done in well- ... In this example protocol, Aequorin-wt and mitochondrial mutated aequorin (Asp119Ala) are used.. ...
Aequorin Studies. Aequorin was loaded into the muscle preparations by the macroinjection technique as previously described.16 ... After aequorin loading and equilibration, muscle preparations were exposed to concentrations of Ca2+ (0.6, 1.2, 2.5, and 5 mmol ... with the bioluminescent indicator aequorin) was prolonged, and abundance of Na+/Ca2+ exchanger mRNA levels increased in ...
Aequorin measurements. The probes used (cytAEQ, erAEQmut, and mtAEQ) are chimeric aequorins targeted to the cytosol, ... The aequorin luminescence data were calibrated off-line into [Ca2+] values, using a computer algorithm based on the Ca2+ ... All aequorin measurements were carried out in KRB supplemented with either 1 mM CaCl2 (cytAEQ and mtAEQ) or 100 μM EGTA ( ... E, cytosolic calcium concentration ([Ca2+]c) by native aequorin with the addition of histamine, [Ca2+]c: mock 1.65 ± 0.13 μM ...
... the crystal structures of calcium-loaded apo-aequorin and apo-obelin. ... Aequorin-1 Chain: A Molecule details › Chain: A. Length: 191 amino acids. Theoretical weight: 21.89 KDa. Source organism: ... All three Ca2+-binding loops of photoproteins bind calcium ions: the crystal structures of calcium-loaded apo-aequorin and apo- ...
0050]Aequorin is taken up by hippocampal neurons. In a set of preliminary studies, aequorin was bilaterally injected directly ... Aequorin is not exported or secreted by cells, nor is it compartmentalized or sequestered within cells. Accordingly, aequorin ... 0016]Aequorin is a photoprotein originally isolated from luminescent jellyfish and other marine organisms. The aequorin complex ... 0019]The function of aequorin is distinguished by several characteristics: aequorin is non-toxic and does not interfere with ...
CHO-K1 Cells expressing GFP-AEQUORIN in Mitochondria and Galpha16, High Quality-Price Ratio Products and Services. Quality ... Home Page > PRIMARY CELL > COMMON USED CELL LINES > CHO-K1 Cells expressing GFP-AEQUORIN in Mitochondria and Ga16 CHO-K1 Cells ... CHO-K1GFP-AEQ cells expressing the mitochondrially-targeted GFP-AEQUORIN protein and Galpha16 in cytoplasma. CHO-K1GFP-AEQ ... cells are first selected from puromycin resistance CHO0K1 cells infected with lentiviruses expressing GFP-AEQUORIN with ...
S1). We then measured the effect of WFS1 loss of function on Ca2+ flux using the ER-targeted aequorin (erAEQ; Fig. 2A) (19). ... Aequorin signals were measured in KRB supplemented with either 1 mM CaCl2 or 100 μM EGTA, using a purpose-built luminometer. ... S.P. and A.D. performed aequorin experiments. J.R. performed in situ PLA and mitochondrial content quantification. C.A.A. and J ... Subcellular calcium measurements in mammalian cells using jellyfish photoprotein aequorin-based probes. Nat. Protoc. 8, 2105- ...
... several red-shifted aequorins, including the most red-shifted aequorin to date, with half-lives of up to 60 s were developed, ... Aequorin was also genetically linked to a VEGFA targeting molecule, a DARPin designated as MP0112, for the imaging of ... Aequorin is a well-characterized bioluminescent photoprotein that has found application in in vitro and in vivo studies. ... In order to increase the applications of aequorin, we have taken established methods that hijack the cellular machinery used to ...
Aequorin Measurements. The aequorin chimera targeted to the ER (erAEQ) (Montero et al. 1995) was transfected alone (control) or ... 1995) Transfected aequorin in the measurement of cytosolic Ca2+ concentration ([Ca2+]c). A critical evaluation. J. Biol. Chem. ... For aequorin measurements, we cotransfected 0.5 μg erAEQ plus 1.5 μg of the construct of interest (or 0.7 μg of each in the ... All aequorin measurements were carried out in KRB and terminated by lysing the cells with 100 μM digitonin in a hypotonic Ca2+- ...
ADP [0.5 microM] produced a rise in [Cai2+] that was registered by both aequorin and quin2 in platelets in Ca2+-containing ... Cai2+] response to A23187 and thrombin was reduced by addition of EGTA to platelets loaded with either aequorin or quin2. With ... With epinephrine, a rise in [Cai2+] was indicated by aequorin, but not by quin2; [Cai2+] signals, aggregation, and secretion ... Platelet activation was better correlated with changes in [Cai2+] indicated by aequorin than with the response of quin2, ...
N2 - An assay was developed for an octapeptide by using recombinant aequorin as the label. Aequorin (AEQ) is a bioluminescent ... AB - An assay was developed for an octapeptide by using recombinant aequorin as the label. Aequorin (AEQ) is a bioluminescent ... An assay was developed for an octapeptide by using recombinant aequorin as the label. Aequorin (AEQ) is a bioluminescent ... abstract = "An assay was developed for an octapeptide by using recombinant aequorin as the label. Aequorin (AEQ) is a ...
2. The aequorin light emission in a medium simulating the in vivo cationic conditions for barnacle muscle fibres indicates that ... 1. The effect of K+, Na+, Mg2+ and pH upon the rate of aequorin utilization has been investigated in the presence of Ca2+. ... 7. The reaction schemes used for the explanation of these and other published results on aequorin (Moisescu, D.G., Ashley, C.C ... 10(-5) M. However, for free calcium concentrations lower than 10(-6) M, the intensity of light emitted by aequorin shows a ...
Aequorin-based homogeneous cortisol immunoassay for analysis of saliva samples. Laura Rowe, Sapna Deo, Josh Shofner, Mark Ensor ... Aequorin-based homogeneous cortisol immunoassay for analysis of saliva samples. / Rowe, Laura; Deo, Sapna; Shofner, Josh; Ensor ... Rowe, L., Deo, S., Shofner, J., Ensor, M., & Daunert, S. (2007). Aequorin-based homogeneous cortisol immunoassay for analysis ... Aequorin-based homogeneous cortisol immunoassay for analysis of saliva samples. Bioconjugate Chemistry. 2007 Nov 1;18(6):1772- ...
Aequorin Loading. Aequorin loading was performed as described previously.6 Briefly, 3 to 5 μL of an aequorin-containing ... Triton X-100 to lyse the aequorin-loaded cells and expose all of the remaining aequorin to Ca2+. This resulted in an ... Aequorin light signals were recorded on a 4-channel recorder in parallel with the LV pressure and coronary perfusion pressure ... The heart was then positioned in a organ bath with the aequorin-loaded area of the LV directed toward the cathode of a ...
Aequorin is not exported or secreted, nor is it compartmentalized or sequestered within cells; thus, aequorin measurements can ... Figure 19.5.8 Images of Ca2+ waves in gastrulating zebrafish embryos detected by microinjected f aequorin (recombinant aequorin ... Recombinant Aequorin. Conventional purification of aequorin from the jellyfish Aequorea victoria requires laborious extraction ... In several experimental systems, aequorins luminescence was detectable many hours to days after cell loading.. Aequorin also ...
mitochondrially targeted aequorin;. PCG.6,. CHO/G16 cell line stably transfected with the Drosophila AKH receptor DNA;. Scg-AKH ...
We performed a structure-activity study with the human motilin receptor, which was recently cloned from thyroid tissue. N-terminal fragments, Ala-analogs of motilin, and motilides were tested in a cell line that expresses the cloned human motilin receptor and apoaequorin. Full potency to induce calc …
The sensitivity of aequorin is due to the fact that bioluminescence is a rare phenomenon in nature and, therefore, it does not ... Aequorin is a non-toxic photoprotein that can be used as a sensitive marker for bioluminescence in vivo imaging. ... This is the first time that aequorin variants incorporating non-canonical amino acids have shown to be active in vivo and ... It is this reaction that endows aequorin with unique characteristics, making it ideally suited for a number of applications in ...
Here we describe a ratiometric low-affinity Ca2+ sensor of the GFP-aequorin protein (GAP) family optimized for measurements in ...
Aequorin Reconstitution and [Ca2+]cytMonitoring. For in vivo aequorin reconstitution (Knight and Knight, 1995), the protoplasts ... but also by the absolute amount of aequorin present in the cells. Higher or lower amounts of aequorin will give larger or ... 1995) Recombinant aequorin methods for intracellular calcium measurement in plants. Methods Cell Biol 49:201-216. ... Influence of db-cGMP (A), db-cAMP (B), 8-Br-cGMP (C), and 8-Br-cAMP (D) on Ca2+-dependent chemiluminescence of aequorin in ...
  • Aequorin is a calcium-activated photoprotein isolated from the hydrozoan Aequorea victoria. (wikipedia.org)
  • The early successful purification of aequorin led to the first experiments involving the injection of the protein into the tissues of living animals to visualize the physiological release of calcium in the muscle fibers of a barnacle. (wikipedia.org)
  • Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium. (nih.gov)
  • Microinjected aequorin has been widely used for intracellular calcium measurement in animal cells, but its use in plants has been limited to exceptionally large cells. (nih.gov)
  • We show here that aequorin can be reconstituted in transformed plants and that it reports calcium changes induced by touch, cold-shock and fungal elicitors. (nih.gov)
  • Compositions containing aequorin and methods for their use in preventing and/or alleviating symptoms and disorders related to calcium imbalance are provided by the present invention. (patentsencyclopedia.com)
  • 4. A method for preventing or alleviating a symptom or disorder associated with calcium imbalance comprising administering to a subject in need of such treatment an effective amount of aequorin. (patentsencyclopedia.com)
  • In particular, this invention is directed to aequorin-containing pharmaceutical and nutraceutical compositions useful in preventing and/or alleviating diseases or symptoms associated with calcium imbalance. (patentsencyclopedia.com)
  • Using ER-targeted aequorin (erAEQ), we have found that S1T proteins reduce ER calcium and reverse elevation of ER calcium loading induced by SERCA1 and SERCA2b. (rupress.org)
  • 2. The aequorin light emission in a medium simulating the in vivo cationic conditions for barnacle muscle fibres indicates that two Ca2+ are apparently involved in this process for free calcium concentrations higher than approx. (edu.au)
  • 10(-5) M. However, for free calcium concentrations lower than 10(-6) M, the intensity of light emitted by aequorin shows a steeper dependency upon [Ca2+] than the square low relationship, indicating that a third Ca2+ should be involved in the process of aequorin light emission, as it has been previously predicted (Moisescu, D.G., Ashley, C.C. and Campbell, A.K. (1975) Biochim. (edu.au)
  • 8. Based on these parameters one can make accurate quantitative predictions for the aequoring light response under a variety of ionic conditions, and this suggests that it is possible to determine absolute free calcium concentrations providing that the ionic composition of the solutions is known, and that the relative rate of aequorin utilization is higher than 0.005. (edu.au)
  • Picosecond fluorescence relaxation spectroscopy of the calcium-discharged photoproteins aequorin and obelin. (wur.nl)
  • Aequorin emits blue light in a calcium dependent manner, while GFP emits green light when irradiated with light of 488 nm, which also led to its name. (news-medical.net)
  • The patent covers the use of aequorin-containing compounds for the purpose of preventing and alleviating symptoms and disorders related to calcium imbalance which include Alzheimer's disease. (bio-medicine.org)
  • Based on our ongoing research of aequorin in various health conditions and what we know about the role of calcium in the body, we expect aequorin to be a vital protein in many aspects of healthy aging. (bio-medicine.org)
  • Comparison of calcium mobilities using Aequorin, Fluo-3, and Fura-2 receptor-expressing cells. (hamamatsu.com)
  • A company called Quincy Bioscience sells a product containing aequorin (Prevagen) that it promotes as "the first supplement to address aging through the restoration of calcium-binding proteins. (thecamreport.com)
  • Aequorin and Photina cell lines - the alternative calcium flux assay. (perkinelmer.com)
  • Calcium-Sensitive Adenylyl Cyclase/Aequorin Chimeras as Sensitive Probes for Discrete Modes of Elevation of Cytosolic Calcium. (elsevier.com)
  • We applied different regimes of temperature changes with well‐defined cooling rates to intact roots of Arabidopsis thaliana expressing the calcium‐indicator, aequorin. (deepdyve.com)
  • Aequorin is presumably encoded in the genome of Aequorea. (wikipedia.org)
  • Rao, BDN, Kemple, MD & Prendergast, FG 1980, ' Proton nuclear magnetic resonance and fluorescence spectroscopic studies of segmental mobility in aequorin and a green fluorescent protein from aequorea forskalea ', Biophysical Journal , vol. 32, no. 1, pp. 630-632. (elsevier.com)
  • In the animals, the protein occurs together with the Green fluorescent protein to produce green light by resonant energy transfer, while aequorin by itself generates blue light. (wikipedia.org)
  • It was also noted during the extraction the animal creates green light due to the presence of the green fluorescent protein, which changes the native blue light of aequorin to green. (wikipedia.org)
  • This has also explained the need for a thiol reagent like beta mercaptoethanol in the regeneration of the protein since such reagents weaken the sulfhydryl bonds between cysteine residues, expediting the regeneration of the aequorin. (wikipedia.org)
  • Chemical characterization of aequorin indicates the protein is somewhat resilient to harsh treatments. (wikipedia.org)
  • In untreated SHR, α-myosin heavy chain (MHC) gene expression and protein were decreased, the Ca 2+ transient (with the bioluminescent indicator aequorin) was prolonged, and abundance of Na + /Ca 2+ exchanger mRNA levels increased in comparison to WKY. (ahajournals.org)
  • CHO-K1GFP-AEQ cells expressing the mitochondrially-targeted GFP-AEQUORIN protein and Galpha16 in cytoplasma. (angioproteomie.com)
  • The plasmid was transformed into Escherichia coli, and the octapeptide-aequorin fusion protein was expressed and purified. (elsevier.com)
  • The properties of the octapeptide-aequorin conjugate were studied, and it was observed that the fusion protein retained the bioluminescence characteristics of native aequorin. (elsevier.com)
  • Here we describe a ratiometric low-affinity Ca2+ sensor of the GFP-aequorin protein (GAP) family optimized for measurements in high-Ca2+ concentration environments. (uva.es)
  • The researchers named this protein aequorin. (princeton.edu)
  • We examined a BL-induced transient increase in cytosolic free Ca 2+ in leaves of transgenic A. thaliana of WT plants, phot1 and phot2 mutants, and phot1 phot2 double mutants expressing the Ca 2+ -sensitive luminescent protein aequorin. (pnas.org)
  • A. thaliana transformants expressing the Ca 2+ -sensitive luminescent protein aequorin were used to show that phot1 regulates a BL-induced transient increase in cytoplasmic Ca 2+ concentration ([Ca 2+ ] c ) in deetiolated seedlings ( 18 ). (pnas.org)
  • We measured [Ca(2+)](cyt) non-invasively using aequorin, and targeted aequorin to the guard cell using a guard cell-specific GAL4-green fluorescent protein enhancer trap line. (nih.gov)
  • aequorin and green fluorescent protein (GFP). (news-medical.net)
  • From this material, they isolated a blue luminescent protein called aequorin and a green fluorescent protein, commonly called GFP. (latimes.com)
  • Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. (abcam.com)
  • Aequorin-based Ca2+ assays represent a new paradigm in drug discovery research for cell-based assays for Ca2+-coupled GPCRs and ion channels. (perkinelmer.com)
  • In order to increase the applications of aequorin, we have taken established methods that hijack the cellular machinery used to synthesize proteins to incorporate non-natural amino acids. (miami.edu)
  • Aequorin has proven to be neuroprotective in pre-clinical studies performed at the University of Wisconsin-Milwaukee and has shown to be effective at improving aspects of cognition such as spatial working memory and executive function in human studies conducted by Quincy Bioscience. (bio-medicine.org)
  • One of the methods of choice (reviewed by Mottheakis and Ohler, 2000) for such measurements is the use of cell lines expressing a GPCR and aequorin, such as described by Sheu et al (1993) or Button et al. (bio-medicine.org)
  • By injecting the same cell suspension into each of the 96 wells, this method avoids the need to wash the dispenser(s) between each measurement and allows 96 measurements of agonist-induced aequorin light emission in 32 minutes with a single-dispenser luminometer. (bio-medicine.org)
  • Aequorin was also genetically linked to a VEGFA targeting molecule, a DARPin designated as MP0112, for the imaging of neovascularization in vivo using a wet age-related macular degeneration model in mice induced by laser exposure. (miami.edu)
  • Aequorin is a non-toxic photoprotein that can be used as a sensitive marker for bioluminescence in vivo imaging. (valpo.edu)
  • This is the first time that aequorin variants incorporating non-canonical amino acids have shown to be active in vivo and useful as reporters in bioluminescence imaging. (valpo.edu)
  • In this study, using Drosophila and taking advantage of an in vivo bioluminescence Ca 2+ -imaging technique in combination with genetic and pharmacological tools, first we show that the GFP-aequorin Ca 2+ sensor is sensitive enough to detect odor-induced responses of various durations. (biologists.org)
  • The UEC group has attempted to use mass spectrometry to elucidate the active center of aequorin bioluminescence, while the OU groups have mainly concerned elucidating the post-translational modification of Phosrestin, which is in fly photoreceptor cells undergoing light-induce reversible phosphorylation in vivo, and determining the in vivo phosphorylation site by mass spectrometry. (nii.ac.jp)
  • Fluoresces in vivo upon receiving energy from the Ca 2+ -activated photoprotein aequorin. (abcam.com)
  • 4. The method of claim 1 wherein said photoprotein labeled reagent comprises a photoprotein selected from the group consisting of aequorin, obelin, mitrocomin, clytin, and combinations thereof. (google.com)
  • A cortisol derivative was chemically conjugated to the lysine residues of a genetically modified aequorin in order to prepare an aequorin-cortisol conjugate capable of binding anticortisol antibodies. (elsevier.com)
  • The binding of anticortisol antibodies to the aequorin-cortisol conjugate resulted in a linear response reflected in the emission of bioluminescence by aequorin. (elsevier.com)
  • A competitive binding assay was developed by simultaneously incubating the aequorin-cortisol conjugate, the anticortisol antibodies, and the sample containing free cortisol. (elsevier.com)
  • 7. The method according to claim 4, wherein said effective amount of aequorin is provided in combination with an immune boosting agent, anti-inflammatory agent, anti-oxidant agent, anti-viral agent, or a mixture thereof. (patentsencyclopedia.com)
  • 8. The method according to claim 4 wherein said effective amount of aequorin is in a unit dosage form selected from the group consisting of a tablet, a capsule, a solution, a suspension, a syrup, a beverage, an oral or ophthalmic formulation and an injection. (patentsencyclopedia.com)
  • 5. The method of claim 4 wherein said photoprotein is aequorin. (google.com)
  • Despite the many advantages of aequorin, its application has been limited by the finite number of canonical amino acids restricting the engineering of aequorin. (miami.edu)
  • Red-Shifted Aequorin Variants Incorporating Non-Canonical Amino Acids:" by Kristen M. Grinstead, Laura Rowe et al. (valpo.edu)
  • The effect of physiologically occurring cations upon aequorin light emission. (edu.au)
  • 3. The inhibitory effect of physiologically occurring cations upon the aequorin light emission can be explained by the cooperative action of two cations, competing with Ca2+ for the reactive sites on aequorin. (edu.au)
  • 5. The experiments indicate a strong interaction between Na+ and K+ in this inhibitory process, since for a given total concentration of monovalent cations, a mixture containing both Na+ and K+ has a larger inhibitory effect on the aequorin light response than solutions containing either Na+ or K+ alone. (edu.au)
  • The L-4- azidophenylalanine substituted aequorin was successfully covalently linked to a fluorophore via the azide to alkyne click reaction for BRET. (miami.edu)
  • 7. The reaction schemes used for the explanation of these and other published results on aequorin (Moisescu, D.G., Ashley, C.C. and Campbell, A.K. (1975) Biochim. (edu.au)
  • It is this reaction that endows aequorin with unique characteristics, making it ideally suited for a number of applications in bioanalysis and imaging. (valpo.edu)
  • An additional non-natural amino acid, L-4-azidophenylalanine, for bio-orthogonal linking via click chemistry was incorporated at position 69 of aequorin. (miami.edu)