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).
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.
Techniques used for determining the values of photometric parameters of light resulting from LUMINESCENCE.
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.
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.
A class of Arthropoda that includes SPIDERS; TICKS; MITES; and SCORPIONS.
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.
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.
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.
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).
Conical muscular projections from the walls of the cardiac ventricles, attached to the cusps of the atrioventricular valves by the chordae tendineae.
An order of MAMMALS, usually flesh eaters with appropriate dentition. Suborders include the terrestrial carnivores Fissipedia, and the aquatic carnivores PINNIPEDIA.
The protein components of a number of complexes, such as enzymes (APOENZYMES), ferritin (APOFERRITINS), or lipoproteins (APOLIPOPROTEINS).
An adrenergic-beta-2 antagonist that has been used for cardiac arrhythmia, angina pectoris, hypertension, glaucoma, and as an antithrombotic.
Compound such as LUMINESCENT PROTEINS that cause or emit light (PHYSICAL LUMINESCENCE).
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.
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.
A benzothaizole which is oxidized by LUCIFERASES, FIREFLY to cause emission of light (LUMINESCENCE).
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.
A chelating agent relatively more specific for calcium and less toxic than EDETIC ACID.
Contractile activity of the MYOCARDIUM.
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.

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+
GFP_AEQVI] Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. ...
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 ...
Microdomains of high intracellular calcium ion concentration, [Ca2+]i, have been hypothesized to occur in living cells exposed to stimuli that generate inositol 1,4,5-trisphosphate (IP3). Mitochondrially targeted recombinant aequorin was used to show that IP3-induced Ca2+ mobilization from intracellular stores caused increases of mitochondrial Ca2+ concentration, [Ca2+]m, the speed and amplitude of which are not accounted for by the relatively small increases in mean [Ca2+]i. A similar response was obtained by the addition of IP3 to permeabilized cells but not by perfusion of cells with Ca2+ at concentrations similar to those measured in intact cells. It is concluded that in vivo, domains of high [Ca2+]i are transiently generated close to IP3-gated channels and sensed by nearby mitochondria; this may provide an efficient mechanism for optimizing mitochondrial activity upon cell stimulation. ...
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 ...
Apoaequorin Reviews - Click here to read this exclusive Apoaequorin review! Does it work? Get the facts. Learn more about this product today!
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 ...
If we are going to be revising the charter, I propose we rename the newsgroup bionet.molbio.proteins.luminescent to cover both photoluminescence (aka fluorescence) and chemiluminescence (bioluminescence). While the present charter stresses bioluminescence, 90% of the discussions have been devoted to GFP and various bioengineered variants. This is not a problem, but it might confuse people looking for bioluminescent proteins. I strongly support some sort of moderation to remove the spams. --John ...
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
Dense core granules of exocrine cells are a potential source of calcium, and results to date have been controversial regarding whether the calcium in these organelles is involved in controlling the cytosolic calcium concentration. Mitchell et al. developed a fusion protein between the dense core transmembrane protein VAMP and the calcium-sensitive bioluminescent protein aequorin in order to visualize calcium in the dense core granules of a pancreatic β cell line. Vesicular free [Ca2+] was approximately 50 μM, significantly lower than endoplasmic reticulum (ER) or Golgi free [Ca2+]. Vesicular [Ca2+] increased when calcium was reintroduced to the cells following calcium depletion. Uptake required adenosine triphosphate (ATP) and was inhibited by high concentrations of orthovanadate, but not by the ER Ca2+ ATPase inhibitor thapsigargin or by agents that altered intravesicular pH or elevations in Na+ concentration, suggesting that the uptake does not occur through a Ca2+/H+ exchanger or through a ...
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.
ASM journals are the most prominent publications in the field, delivering up-to-date and authoritative coverage of both basic and clinical microbiology.. About ASM , Contact Us , Press Room. ASM is a member of. ...
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:. ...
Female rats were fed for 21 days on 5 semi-synthetic diets containing 8 or 30% proteins, 1 or 25% lipids respectively, the… Expand ...
If you have a ferret, then this article is for you! We answer 10 FAQs about when do ferrets shed. How much do they shed? Why does it happen? And more
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 derive from the same animal. Apoaequorin, the protein portion of aequorin, is an ...
Other cofactors may be required, such as calcium (Ca2+) for the photoprotein aequorin, or magnesium (Mg2+) ions and ATP for the ... Furthermore, some of the blue light released by aequorin in contact with calcium ions is absorbed by a green fluorescent ... 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 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- ... In 1961, Shimomura and Johnson isolated the protein aequorin, and its small molecule cofactor, coelenterazine, from large ...
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. Bibcode:1975Natur.256 ... 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 ...
Borle, A (1986). "A Simple Method for Incorporating Aequorin into Mammalian-Cells". American Journal of Physiology. 251 (2): ...
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 ...
Feeds on brine shrimp (Artemia salina) and rotifers (Brachionus plicatilis). It is bioluminescent due to aequorin and green ...
Colocalization of aequorin with GFP facilitates BRET/CRET (Bioluminescence or Chemiluminescence Resonance Energy Transfer), ... Such systems may rely on aequorin and the luciferin coelenterazine. Ca2+ binding causes a conformational change that ... "Chimeric green fluorescent protein-aequorin as bioluminescent Ca2+ reporters at the single-cell level". Proceedings of the ...
George CH, Kendall JM, Campbell AK, Evans WH (November 1998). "Connexin-aequorin chimerae report cytoplasmic calcium ... "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 ...
GFP is co-expressed with aequorin in small granules around the rim of the jellyfish bell. The secondary excitation peak (480 nm ... In the 1960s and 1970s, GFP, along with the separate luminescent protein aequorin (an enzyme that catalyzes the breakdown of ... In A. victoria, GFP fluorescence occurs when aequorin interacts with Ca2+ ions, inducing a blue glow. Some of this luminescent ... The purpose of both the (primary) bioluminescence (from aequorin's action on luciferin) and the (secondary) fluorescence of GFP ...
... along with the separate luminescent protein aequorin (an enzyme that catalyzes the breakdown of luciferin, releasing light), ... purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea". Journal of ...
Shimomura, O.; Johnson, F. H.; Saiga, Y. (1962). "Extraction, purification and properties of aequorin, a bioluminescent protein ... In 1961, Osamu Shimomura extracted green fluorescent protein (GFP) and another bioluminescent protein, called aequorin, from ...
Prendergast FG, Mann KG (August 1978). "Chemical and physical properties of aequorin and the green fluorescent protein isolated ...
Despite containing a bioluminescent protein, aequorin, this species (as well as all other species in the genus) are almost ... Prendergast, Franklyn G.; Mann, Kenneth G. (1978-08-22). "Chemical and physical properties of aequorin and the green ... and extracted aequorin and GFP from various Aequorea species including A. forskalea. To this day, A. victoria is still the ... aequorin) and GFP (green fluorescent protein) discovered and studied extensively by Dr. Osamu Shimomura in 1961. While this ...
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. (8 August 1991). "Transgenic plant aequorin ... 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 cnidarian ...
"Determination of affinity and activity of ligands at the human neuropeptide Y Y4 receptor by flow cytometry and aequorin ...
In 1962, their work culminated in the discovery of the proteins aequorin and green fluorescent protein (GFP) in A. victoria; ...
This dragon is under the permanent effect of a Silhouette Scroll. A toggle on the dragons profile allows swapping between the artwork poses available for the breed ...
T2 - Effects of OR-1896, an active metabolite of levosimendan, on contractile force and aequorin light transients in intact ... Erratum: Effects of OR-1896, an active metabolite of levosimendan, on contractile force and aequorin light transients in intact ... title = "Erratum: Effects of OR-1896, an active metabolite of levosimendan, on contractile force and aequorin light transients ... Erratum : Effects of OR-1896, an active metabolite of levosimendan, on contractile force and aequorin light transients in ...
The recombinant aequorin gene from the jellyfish Aequorea victoria responsible for the expression of the Ca2+-sensitive ... aequorin photoprotein has been cloned in the filamentous fungus Aspergillus awamori. This has allowed real life monitoring of [ ... bioassay utilizing eukaryotic cells has been developed based on filamentous fungi transformed with the recombinant aequorin ... Recently recombinant aequorin gene has been expressed in filamentous fungi [10]. Aequorin is a Ca2+-sensitive photoprotein of ...
ANTICA TERRA Chardonnay, Willamette Valley Aequorin 2019 The scintillating 2019 Aequorin Chardonnay from winemaker Maggie ...
And when Ca2+ ions bind on aequorin, the aequorin will emit a glowing blue light and GFP will raise to excited state and emit ... Aequorin and GFP(Green Fluorescent Protein)has existed for more than one hundred millions years and were discovered only in one ... Aequorin is composed of two distinct units: a) the apoprotein: apoaequorin and b) the prosthetic group: coelenterazine (a ... In our research we use Aequorin-GFP fusion protein to be strong enough to illuminat and to replace streetlamp and hope one day ...
INTERCHIM AEQUORIN EXPRESSION VECTOR PUC19 VECTOR P/N : BX1020 Pack : 1 x 25 ug ...
He found that calcium ions activate a jellyfish protein called aequorin. Once activated, aequorin produces blue light, but if ... Cloning and expression of the Cdna coding for aequorin, a bioluminescent calcium-binding protein. Biochem Biophys Res Comm 1985 ... Shimomura O, Johnson FH, Y Saiga Y. Extraction, purification and properties of aequorin, a bioluminescent protein from the ...
Targeting transgenic aequorin to root pericycle cells. Trewavas, A.. EU government bodies ...
... and aequorin. This construct does in principle monitor levels of [Ca2+]i close to ion channel microdomains and, provide ... already transformed with firefly luciferase and transfected with the photoprotein aequorin, enabling changes in cytosolic free ... is one of the methods routinely used by cell biologists to measure free Ca2+ inside cells and in particular aequorin a ...
A toolset of aequorin expression vectors for in planta studies of subcellular calcium concentrations in Arabidopsis thaliana. ... cellular response by temporal and spatial changes in cellular calcium concentration and the calcium-sensitive protein aequorin ...
The FLIPR Tetra System is now available with an aequorin option which includes a novel ICCD camera technology optimized for use ... A preconfigured aequorin assay protocol for the FlexStation 3 reader is available in the SoftMax Pro Software. ... The FlexStation 3 reader and the FLIPR Tetra System with aequorin option (ICCD camera) have been used to measure the Photina ... This application note provides a basic protocol for performing an adherent aequorin assay using the FlexStation 3 reader and ...
Regeneration of Aequorin in Aequorea Aequorea jellyfish - http://www.nature.com/nature/journal/v256/n5514/pdf/256236a0.pdf ...
Knight MR, Campbell AK, Smith SM, Trewavas AJ, Recombinant aequorin as a probe for cytosolic free Ca 2+ in Escherichia coli, ... Knight MR, Campbell AK, Smith SM, Trewavas AJ, Transgenic plant aequorin reports the effects of touch and cold-shock and ...
Likewise, Aequorea victoria green fluorescent protein is thought to participate in a tetrameric complex with aequorin, but this ... researchers determined that aequorin and the green fluorescent protein work together in the light organs of the jellyfish to ... which they named aequorin. During the isolation procedure, a second protein was observed that lacked the blue-emitting ... bioluminescent properties of aequorin, but was able to produce green fluorescence when illuminated with ultraviolet light. Due ...
Agonist activity at MmOT receptor expressed in HEK293FT cells assessed as stimulation of calcium release by Aequorin based ...
... are soon place magnetic aequorin like review or Social Security documents. If you are any Roses with the view Физика,, limit ...
This will be achieved by generating peroxisomal-targeted calcium reporters based on aequorin and fluorescent probes which we ... aequorin and fluorescent) which we have utilised in previously published studies. Achieved - Localisation studies with tagged ...
FP with properties comparable to the photoprotein aequorin, and this association ultimately led to cloning the cDNA that ... to its high quantum yield and extinction provigil insomnia coefficient to be a superior energy transfer acceptor for aequorin. ...
... spectra for AvicFP2 and AvicFP3 were measured using a hand-held net and was transported back to the photoprotein aequorin than ...
... and aequorins. It is commonly used to monitor reporter genes in BRET (Bioluminescence Resonance Energy Transfer), ELISA and HTS ... and aequorins. It is commonly used to monitor reporter genes in BRET (Bioluminescence Resonance Energy Transfer), ELISA and HTS ... and aequorins. It is commonly used to monitor reporter genes in BRET (Bioluminescence Resonance Energy Transfer), ELISA and HTS ... and aequorins. It is commonly used to monitor reporter genes in BRET (Bioluminescence Resonance Energy Transfer), ELISA and HTS ...
This study utilized the bioluminescent Ca2+-indicator GFP-aequorin to monitor the nicotine-induced Ca2+-response within the ...
... then they are all likely to be a superior energy transfer acceptor for buy cheap oxytrol online aequorin. While searching for ... GFP continue to inspire us and to the photoprotein aequorin, and this association ultimately led to cloning the cDNA that ...
Bioluminescence inmunoassay for cortisol using recombinant aequorin as a label. Analytical Biochemistry, 306(2), 204-211. Tente ...
... experiments including the utilization of transgenic flowers carrying an inducible gene for aequorin connected to GFP including ...
  • The recombinant aequorin gene from the jellyfish Aequorea victoria responsible for the expression of the Ca 2+ -sensitive aequorin photoprotein has been cloned in the filamentous fungus Aspergillus awamori . (biomedcentral.com)
  • Aequorin and GFP(Green Fluorescent Protein)has existed for more than one hundred millions years and were discovered only in one jellyfish species-Aequorea Victoria. (igem.org)
  • He found that calcium ions activate a jellyfish protein called aequorin. (lww.com)
  • Osamu Shimomura and Frank Johnson, working at the Friday Harbor Laboratories of the University of Washington in 1961, first isolated a calcium-dependent bioluminescent protein from the Aequorea victoria jellyfish, which they named aequorin . (microscopyu.com)
  • Over the next two decades, researchers determined that aequorin and the green fluorescent protein work together in the light organs of the jellyfish to convert calcium-induced luminescent signals into the green fluorescence characteristic of the species. (microscopyu.com)
  • In A. victoria , aequorin emits blue light which excites GFP giving the overall green glow to the jellyfish 1 . (biologists.com)
  • A novel toxicity bioassay utilizing eukaryotic cells has been developed based on filamentous fungi transformed with the recombinant aequorin gene. (biomedcentral.com)
  • Bioluminescence inmunoassay for cortisol using recombinant aequorin as a label. (bvsalud.org)
  • Aequorin photoprotein (untagged) is recombinantly produced in E. coli, purified via multi-step chromatography and appears as a yellow liquid. (nanolight.com)
  • Aequorin photoprotein (untagged) is recombinantly produced in E. coli, purified via multi-step chromatography and appears as a white lyophilized powder. (nanolight.com)
  • b) [Ca2+]c measurement with fura-2 followed by quantitative immunocytochemistry of single cells and iii) [Ca2+]c measurement of cell population upon cotransfection with the Ca(2+)-sensitive photoprotein, aequorin. (rupress.org)
  • During the isolation procedure, a second protein was observed that lacked the blue-emitting bioluminescent properties of aequorin, but was able to produce green fluorescence when illuminated with ultraviolet light. (microscopyu.com)
  • This study utilized the bioluminescent Ca2+-indicator GFP-aequorin to monitor the nicotine-induced Ca2+-response within the mushroom bodies (MB) , a higher-order brain center in flies, and examined how DopEcR modulates these Ca2+-dynamics. (sdbonline.org)
  • Once Ca 2+ ions are bound to the three Ca 2+ -binding sites in aequorin, the protein is converted into an oxygenase. (biomedcentral.com)
  • In our research we use Aequorin-GFP fusion protein to be strong enough to illuminat and to replace streetlamp and hope one day it can substitute daylight lamp without use of electricity. (igem.org)
  • Once activated, aequorin produces blue light, but if the green fluorescent protein is nearby, the two proteins together yield a bright green signal. (lww.com)
  • At that time, they called this protein aequorin. (coursera.org)
  • This distinct discovery happened in a very interesting chance and then, because he was studying the how to make this protein Aequorin to have light, at that time, and then he tried so many solutions and the combinations and he failed. (coursera.org)
  • But one day, when he was about to finish his experiment, he throw out his solutions into the sink, and then, of course, his left over Aequorin, this protein into sink as well, so all of a sudden, he saw in the sink the protein has very strong light over there. (coursera.org)
  • And then the first time, he realized this protein, Aequorin here, might be very useful for biological studies and research. (coursera.org)
  • Two fluorescent proteins were identified in this study: the first, named aequorin, exhibited a blue glow in the presence of Ca 2+ ions. (biologists.com)
  • Then Aequorin can emit the blue light (wavelenght= 469 nm) and supply energy for GFP. (igem.org)
  • Aequorin was "charged" with unmodified (native) Coelenterazine and will emit light at 465 nm upon Ca 2+ contact. (nanolight.com)
  • Description: Agonist activity at MmOT receptor expressed in HEK293FT cells assessed as stimulation of calcium release by Aequorin based assay. (guidetopharmacology.org)
  • Aequorin is composed of two distinct units: a) the apoprotein: apoaequorin and b) the prosthetic group: coelenterazine (a luciferin). (igem.org)
  • Polysciences) was used in calculation of the interactions between AvicFP1 and aequorin are beyond the scope of this species in the weak dimer interface in the. (gotonextstep.com)
  • Endoh, M. / Erratum : Effects of OR-1896, an active metabolite of levosimendan, on contractile force and aequorin light transients in intact rabbit ventricular myocardium (Journal of Cardiovascular Pharmacology (2000) 36 (118-125)) . (elsevier.com)
  • The review will publish desired to high change aequorin. (vanpanhuys.com)
  • For your download Batı'ya Yön Veren Metinler- III Aydınlanma Burjuvazi Yüzyılı Bilim Çağının Zaferi (1650- 1800) 2010 , are soon place magnetic aequorin like review or Social Security documents. (coldbacon.com)