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)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 holoprotein composed of two distinct units, the apoprotein that is called apoaequorin, which has an approximate molecular weight of 21 kDa, and the prosthetic group coelenterazine, the luciferin. (wikipedia.org)
- When coelenterazine is bound, it is called aequorin. (wikipedia.org)
- The crystal structure revealed that aequorin binds coelenterazine and oxygen in the form of a peroxide, coelenterazine-2-hydroperoxide. (wikipedia.org)
- It was later discovered that the apoprotein can stably bind coelenterazine and oxygen is required for the regeneration to the active form of aequorin. (wikipedia.org)
- Aequorin is a calcium-sensitive luminescent protein from the coelenterate Aequorea victoria (A. forskalea) which is formed from apoaequorin, a polypeptide of relative molecular mass approximately 22,000, and coelenterazine, a hydrophobic luminophore. (nih.gov)
- 1993). In this system, cells are incubated with coelenterazine, which is the co-factor of aequorin. (bio-medicine.org)
- During this incubation, coelenterazine enters the cell (it is lipophylic and readily crosses the cell membrane) and conjugates with apoaequorin to form aequorin, which is the active form of the enzyme. (bio-medicine.org)
- This increase leads to the activation of the catalytic activity of aequorin, which oxidizes coelenterazine and yields apoaequorin, coelenteramide, CO 2 and light. (bio-medicine.org)
- Cells expressing apoaequorin and a GPCR are detached from the culture plate (if they are adherent) and are incubated with coelenterazine to reconstitute active aequorin. (bio-medicine.org)
- Reconstitution of an active recombinant aequorin in living cells is obtained by simple addition of coelenterazine into the medium. (bio-protocol.org)
- By site-specifically incorporating the non-natural amino acids L-4-aminophenylalanine, L-4-bromophenylalanine, L-4-iodophenylalanine, and L-4-methoxyphenylalanine, into positions associated with the bioluminescence and charging them with analogs of coelenterazine, several red-shifted aequorins, including the most red-shifted aequorin to date, with half-lives of up to 60 s were developed, creating aequorin mutants suitable for multiplexing and for transparent and deep tissue imaging. (miami.edu)
- Emission of bioluminescence in the blue-region of the spectrum by aequorin only occurs when calcium, and its luciferin coelenterazine, are bound to the protein and trigger a biochemical reaction that results in light generation. (valpo.edu)
- Herein we report the site-specific incorporation of non-canonical or non-natural amino acids and several coelenterazine analogues, resulting in a catalog of 72 cysteine-free, aequorin variants which expand the potential applications of these photoproteins by providing several red-shifted mutants better suited to use in vivo . (valpo.edu)
- In vivo studies in mouse models using the transparent tissue of the eye confirmed the activity of the aequorin variants incorporating L-4-iodophehylalanine and L-4-methoxyphenylalanine after injection into the eye and topical addition of coelenterazine. (valpo.edu)
- Squid use coelenterazine and a luciferase called aequorin . (everything2.com)
- Upon GPCR stimulation and subsequent calcium binding to the aequorin oxidation of coelenterazine leads to emission of light. (perkinelmer.com)
- The apo-enzyme (apoaequorin) is a 21 kD protein, which requires a hydrophobic prosthetic group, coelenterazine, to be converted to aequorin,the active form of the enzyme. (perkinelmer.com)
- When activated by calcium in the cell, this enzyme, called aequorin, reacts with another jellyfish compound called coelenterazine. (technologyreview.com)
- Discussions of "jellyfish DNA" to make "glowing" animals often refer to transgenic animals which express the Green fluorescent protein, not aequorin, although both originally derived from the same animal. (wikipedia.org)
- Aequorin is a 21 kDa photoprotein isolated from jellyfish Aequorea victoria that emits blue light in the presence of Ca 2+ . (bio-protocol.org)
- Aequorin (AEQ) is a bioluminescent protein found in the jellyfish (Aequorea victoria). (elsevier.com)
- aequorin pairing that is used in jellyfish. (everything2.com)
- In the 1960s and 1970s, GFP, along with the separate luminescent protein aequorin (an enzyme that catalyzes the breakdown of luciferin , releasing light), was first purified from the jellyfish Aequorea victoria and its properties studied by Osamu Shimomura . (wikipedia.org)
- Aequorin, which comes from a species of jellyfish called Aequorea victoria, is a calcium-binding protein. (bio-medicine.org)
- A reader asked what I've heard about this product, which contains aequorin (Aequorea victoria), a calcium-binding protein found in the jellyfish. (thecamreport.com)
- Aequorin is a photo protein originating from the jellyfish Aequorea Victoria. (perkinelmer.com)
- 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)
- Cell lines expressing recombinant aequorin and a G-protein coupled receptor for functional screening ( INTRODUCTION Man. (bio-medicine.org)
- Thanks to recombinant cDNAs expression, aequorin can be specifically targeted to various subcellular compartments, thus allowing an accurate measurement of Ca 2+ uptake and release of different intracellular organelles. (bio-protocol.org)
- Aequorin began to be widely used when the cDNA encoding the photoprotein was cloned, thus opening the way to recombinant expression. (bio-protocol.org)
- An assay was developed for an octapeptide by using recombinant aequorin as the label. (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)
- Since the emitted light can be easily detected with a luminometer, aequorin has become a useful tool in molecular biology for the measurement of intracellular Ca2+ levels. (wikipedia.org)
- Aequorin is a Ca 2+ sensitive photoprotein suitable to measure intracellular Ca 2+ transients in mammalian cells. (bio-protocol.org)
- 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)
- Aequorin is a well-characterized bioluminescent photoprotein that has found application in in vitro and in vivo studies. (miami.edu)
- In this article, we describe the development of a homogeneous assay for the hormone cortisol using the bioluminescent photoprotein aequorin as a reporter molecule. (elsevier.com)
- In the aequorin assay, cells co-expressing apoaequorin and the target receptor are first incubated with the co-factor coelenterazinein order to reconstitute the activea equorin enzyme. (perkinelmer.com)
- In A. victoria , GFP fluorescence occurs when aequorin interacts with Ca 2+ ions, inducing a blue glow. (wikipedia.org)
- Aequorin is the luciferase used by other marine animals, like squid, but GFP is not a luciferase. (everything2.com)
- 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)
- The aequorin transients were abbreviated and demonstrated decreased amplitude. (ahajournals.org)
- 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)
- CHO-K1GFP-AEQ cells are first selected from puromycin resistance CHO0K1 cells infected with lentiviruses expressing GFP-AEQUORIN with mitochondria targeting signal sequence. (angioproteomie.com)