Abstract:
:Cellular calcium signaling events are transient. Hence they are observed in real time using fluorescence imaging or electrophysiological methods that require sophisticated instrumentation and specialized skills. For high throughput assays simple and inexpensive techniques are desirable. Many calcium channels that serve as drug targets have subtypes arising from diverse subunit combinations. These need to be targeted selectively for achieving efficacy and for avoiding side effects in therapies. This in turn increases the number of calcium channels that act as drug targets. We report a novel method for intracellular calcium sensing that utilizes the calcium dependent stable interaction between CaM kinase II (CaMKII) and its ligands such as the NMDA receptor subunit GluN2B. The CaMKII-GluN2B complex formed persists as a memory of the transient increase in calcium. In a cell-based assay system GFP-α-CaMKII expressed in the cytosol responds to calcium by translocating towards GluN2B sequence motif exogenously expressed on mitochondria or endoplasmic reticulum. The resulting punctate fluorescence pattern serves as the signal for intracellular calcium release. The pattern is stable, unaffected by sample processing and is observable without real time imaging. The activities of calcium channel proteins heterologously expressed in HEK-293 cells were detected with specificity using this technique. A calcium sensor vector and a calcium sensor cell line were developed as tools to perform this technique. This technique being simple and less expensive could significantly facilitate high throughput screening in calcium channel drug discovery.
journal_name
Cell Calciumjournal_title
Cell calciumauthors
Chandrika AR,Steephan M,Raveendran Nair R,Sudarsana Devi SP,Kumar M,Paul S,Madhavan M,Omkumar RVdoi
10.1016/j.ceca.2018.05.009subject
Has Abstractpub_date
2018-09-01 00:00:00pages
73-85eissn
0143-4160issn
1532-1991pii
S0143-4160(17)30170-7journal_volume
74pub_type
杂志文章相关文献
CELL CALCIUM文献大全abstract::Pancreatic islets produce pulses of insulin and other hormones that maintain normal glucose homeostasis. These micro-organs possess exquisite glucose-sensing capabilities, allowing for precise changes in pulsatile insulin secretion in response to small changes in glucose. When communication among these cells is disrup...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2019.102081
更新日期:2019-11-01 00:00:00
abstract::STIM and Orai isoforms orchestrate store operated Ca2+ entry (SOCE) and thus cytosolic Ca2+ fluctuations following stimulation by hormones, growth factors and further mediators. Orai1 is a target of Nedd4-2, an ubiquitin ligase preparing several plasma membrane proteins for degradation. Phosphorylation of Nedd4-2 by t...
journal_title:Cell calcium
pub_type: 杂志文章,评审
doi:10.1016/j.ceca.2012.05.005
更新日期:2012-11-01 00:00:00
abstract::We studied the contribution of store-operated or capacitative Ca2+-entry (SOCE or CCE, respectively) through store-operated Ca2+ channels (SOCCs) and the contribution of Ca2+-entry through receptor-operated, non-selective cation channels (ROCCs or NSCCs, respectively), on the M3-receptor-mediated (270 nM Ach) contract...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2005.06.035
更新日期:2005-11-01 00:00:00
abstract::In the presence of inositol (1,4,5)-trisphosphate (InsP3) repetitive waves of elevated cytosolic free Ca2+ (Ca waves) that travel through cellular cytoplasm are observed. Investigation of this phenomenon stimulated the view of cellular cytoplasm as 'an excitable medium composed of Ca release processes (InsP3R), couple...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(94)90019-1
更新日期:1994-09-01 00:00:00
abstract::The mechanism by which the depletion of intracellular Ca2+ stores stimulates Ca2+ influx is poorly understood. However, the coupling of depletion to influx is broken during mitosis [Preston, S.F. et. al., (1991) Cell Regul., 2, 915-925]. Thus, in interphase HeLa cells, activation of the histamine H1 receptor, or incub...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(93)90042-5
更新日期:1993-05-01 00:00:00
abstract::A potential role of arachidonic acid in the modulation of insulin secretion was investigated by measuring its effects on calmodulin-dependent protein kinase and protein kinase C in islet subcellular fractions. The results were interpreted in the light of arachidonic acid effects on insulin secretion from intact islets...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(92)90044-s
更新日期:1992-03-01 00:00:00
abstract::A new method for the determination of tissue concentrations of Fura-2 and Fura-2/AM was developed based upon acetonitrile extraction followed by RP-HPLC separation (using tetrahexylammonium as counter-ion), post-column alkaline hydrolysis of Fura-2/AM, and fluorimetric detection. The detection limit was 1.2 nM and 1 n...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(95)90057-8
更新日期:1995-11-01 00:00:00
abstract::Isolated avian growth plate chondrocytes convert the acetoxymethyl ester (AM) form of Fura-2 quickly and efficiently to the Ca2(+)-sensitive pentacarboxylic acid (FA) form. Control experiments indicate that the Kd for intracellular Fura-2/FA is very close to that of extracellular Fura-2/FA at the same ionic strength a...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(90)90077-8
更新日期:1990-08-01 00:00:00
abstract::Many fluorescent Ca indicators, particularly those loaded as acetoxymethyl (AM)-ester derivatives, are known to become compartmentalized into organelles. This property can be exploited to measure changes in free [Ca] in subcellular compartments, including the inositol (1,4,5)-trisphosphate-sensitive store. However, qu...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/s0143-4160(96)90029-9
更新日期:1996-09-01 00:00:00
abstract::The sensitivity to Ca2+ of the Ca2+-dependent K+ channel can be increased by the artificial electron donor system ascorbate + phenazine-methosulphate in a variety of animal cells. In the human erythrocyte the shift from the 'low' to the 'high-affinity' state seems to depend on the reduction of a membrane component acc...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(83)90024-6
更新日期:1983-12-01 00:00:00
abstract::The high capacity, low affinity Ca(2+) storage protein chromogranins are marker proteins of secretory granules that contain the most Ca(2+) in secretory cells. Along with the abundantly expressed chromogranins, the IP(3)R/Ca(2+) channels, the major intracellular Ca(2+) channels, are also expressed in secretory granule...
journal_title:Cell calcium
pub_type: 杂志文章,评审
doi:10.1016/j.ceca.2011.12.008
更新日期:2012-03-01 00:00:00
abstract::Ca2+ efflux from dog red blood cells loaded with Ca2+ using the A23187 ionophore could be separated into two main components: (1) Mg- and ATP-dependent (active transport) and (2) dependent on external Na (K1/2 around 15 mM); at 80 microM internal free Ca the relative magnitudes of these fluxes were 70% and 30% respect...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(85)90025-9
更新日期:1985-12-01 00:00:00
abstract::The surface dynamics of neurotransmitter receptors and transporters, as well as ion channels, has been well-documented in neurons, revealing complex molecular behaviour and key physiological functions. However, our understanding of the membrane trafficking and dynamics of the signalling molecules located at the plasma...
journal_title:Cell calcium
pub_type: 杂志文章,评审
doi:10.1016/j.ceca.2017.08.009
更新日期:2017-11-01 00:00:00
abstract::Fluorescence imaging of calcium ions (Ca(2+)) has become an essential technique for investigation of signaling pathways involving Ca(2+) as a second messenger. But, Ca(2+) signaling is involved in many biological phenomena, and therefore simultaneous visualization of Ca(2+) and other biomolecules (multicolor imaging) ...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2016.06.002
更新日期:2016-10-01 00:00:00
abstract::Here we report on the feasibility of using replication deficient adenoviruses to modify signal transduction systems in epithelia. We constructed two viruses, one expressing a dominant negative mutant of the alpha-subunit of Gq (Ad-EF1-dnG alpha q) and the other expressing the wild-type alpha-subunit of Gq (Ad-EF1-wtG ...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/s0143-4160(98)90077-x
更新日期:1998-08-01 00:00:00
abstract::Here, we report a novel role for the cysteine protease calpain in store-operated calcium entry. Several structurally and mechanistically unrelated inhibitors of calpain inhibited Ca2+ entry activated in human platelets by thapsigargin-evoked Ca2+ store depletion or the physiological agonist thrombin, whereas inhibitor...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2006.05.008
更新日期:2007-02-01 00:00:00
abstract::Antisense oligodeoxynucleotides (AS-ODNs) were used in combination with transient functional expression of the cardiac Na(+)-Ca2+ exchanger (NCX1) to correlate suppression of the Na(+)-Ca2+ exchange function with down-regulation of NCX1 protein expression. In a de-novo expression system (Sf9 cells), a decrease in both...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2004.10.006
更新日期:2005-03-01 00:00:00
abstract::Specific sequences (cis-acting elements) in the 3'-untranslated region (UTR) of RNA, together with stabilizing and destabilizing proteins (trans-acting factors), determine the mRNA stability, and consequently, the level of expression of several proteins. Such interactions were discovered initially for short-lived mRNA...
journal_title:Cell calcium
pub_type: 杂志文章,评审
doi:10.1016/j.ceca.2006.04.004
更新日期:2006-10-01 00:00:00
abstract::Signaling between intracellular Ca2+ stores and cell membrane channels or transporters is important to Ca(2+)-based second messenger systems. Two hypotheses, the capacitative and the Ca(2+)-induced Ca(2+)-influx models have been proposed to explain aspects of this signaling. In this study, we examined the applicabilit...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(94)90076-0
更新日期:1994-12-01 00:00:00
abstract::A characteristic feature of developing neural networks is spontaneous periodic activity. In the developing retina, retinal ganglion cells fire bursts of action potentials that drive large increases in intracellular calcium concentration with a periodicity of minutes. These periodic bursts of action potentials propagat...
journal_title:Cell calcium
pub_type: 杂志文章,评审
doi:10.1016/j.ceca.2005.01.010
更新日期:2005-05-01 00:00:00
abstract::NMDA receptors (NMDARs) are glutamate-gated ion channels involved in excitatory synaptic transmission and in others physiological processes such as synaptic plasticity and development. The overload of Ca2+ ions through NMDARs, caused by an excessive activation of receptors, leads to excitotoxic neuronal cell death. Fo...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2019.04.001
更新日期:2019-06-01 00:00:00
abstract::An inward current responsible for hormone regulated Ca2+ entry has been identified in cultured rat hepatocytes using whole cell patch clamp. Addition of 20 nM vasopressin or of 100 microM ATP induced the inward current, which could be observed more clearly after blocking an outward K+ current. This large outward K+ cu...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(95)90042-x
更新日期:1995-07-01 00:00:00
abstract::The best known calcium waves move at about 5-30 microns/s (at 20 degrees C) and will be called fast waves to distinguish them from slow (contractile) ones which move at 0.1-1 microns/s as well as electrically propagated, ultrafast ones. Fast waves move deep within cells and seem to underlie most calcium signals. Their...
journal_title:Cell calcium
pub_type: 杂志文章,评审
doi:10.1016/0143-4160(93)90099-r
更新日期:1993-11-01 00:00:00
abstract::There is growing evidence indicating that the pore structure of voltage-gated ion channels (VGICs) influences gating besides their conductance. Regarding low voltage-activated (LVA) Ca2+ channels, it has been demonstrated that substitutions of the pore aspartate (D) by a glutamate (D-to-E substitution) in domains III ...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2020.102214
更新日期:2020-07-01 00:00:00
abstract::The importance of Ca2+-signaling for many subcellular processes is well established in higher eukaryotes, whereas information about protozoa is restricted. Recent genome analyses have stimulated such work also with Alveolates, such as ciliates (Paramecium, Tetrahymena) and their pathogenic close relatives, the Apicomp...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2012.01.006
更新日期:2012-05-01 00:00:00
abstract::Rat liver was homogenized and subjected to differential centrifugation. When the low speed nuclear pellet was processed on a Percoll gradient, plasma membrane markers and Ins(1,4,5)P3 binding activity purified together. The high speed (microsomal) fraction was subfractionated by sucrose density gradient centrifugation...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/0143-4160(93)90007-s
更新日期:1993-06-01 00:00:00
abstract::The lungs as the gateways of our body to the external environment are essential for gas exchange. They are also exposed to toxicants from two sides, the airways and the vasculature. Apart from naturally produced toxic agents, millions of human made chemicals were produced since the beginning of the industrial revoluti...
journal_title:Cell calcium
pub_type: 杂志文章,评审
doi:10.1016/j.ceca.2017.04.005
更新日期:2017-11-01 00:00:00
abstract::The strength of the heart beat depends on the amplitude and time course of the transient increase in [Ca(2+)] in the myocytes with each cycle. [Na(+)](i) modulates cardiac contraction through its effect on the Ca(2+) flux through the Na/Ca exchanger. Cardiac excitation-contraction coupling has been postulated to occur...
journal_title:Cell calcium
pub_type: 杂志文章,评审
doi:10.1016/j.ceca.2004.01.014
更新日期:2004-06-01 00:00:00
abstract::Parvalbumin (PA) is a Ca(2+)-binding protein of vertebrates massively expressed in tissues with high oxygen uptake and respectively elevated level of reactive oxygen species (ROS). To characterize antioxidant properties of PA, antioxidant capacity (AOC) of intact rat α-PA has been explored. ORAC, TEAC and hydrogen per...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2014.03.001
更新日期:2014-05-01 00:00:00
abstract::NAADP has been shown to act as a second messenger in a wide range of systems from plants to mammalian cells. Although it had always been considered as a canonical second messenger, recent work has shown that it is also active when applied extracellularly. It has also been suggested that NAADP might have a direct actio...
journal_title:Cell calcium
pub_type: 杂志文章
doi:10.1016/j.ceca.2006.10.002
更新日期:2007-06-01 00:00:00