Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:1.13.12.5 (aequorin)
 1,451 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The cardiac type Na(+)/Ca(2+) exchanger (NCX1) has been transiently expressed in Chinese hamster ovary cells, which do not contain an endogenous exchanger, together with aequorin chimeras that are targeted to different intracellular compartments to investigate intracellular Ca(2+) homeostasis. The expression of NCX decreased the endoplasmic reticulum Ca(2+) concentration, [Ca(2+)](er), in resting cells, showing that the exchanger was operative under these conditions. It induced a greater reduction in the height of the mitochondrial and cytosolic Ca(2+) transients in agonist-stimulated cells than would have been expected from the [Ca(2+)](er) decrease. It also had a major effect on the sub-plasma membrane Ca(2+) concentration, [Ca(2+)](pm): after a transient [Ca(2+)](pm) rise induced by the activation of capacitative Ca(2+) influx, [Ca(2+)](pm) settled to a value about 3-fold higher than in controls. The sustained [Ca(2+)](pm) increase after the transient was due to the operation of the exchanger, either directly by operating in the Ca(2+) entry mode, or indirectly by removing the Ca(2+) inhibition on the capacitative Ca(2+) influx channels.
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PMID:Recombinant expression of the plasma membrane Na(+)/Ca(2+) exchanger affects local and global Ca(2+) homeostasis in Chinese hamster ovary cells. 1214 98

The Na/Ca exchanger is a large-capacity system that is supposed to be predominant in the ejection of calcium in excitable tissues. Its activity is normally evaluated in electro-physiological experiments or using Na-loaded cells or plasma membrane vesicles in experiments in which the activity is estimated by measuring the uptake of isotopic calcium. Here the activity of the exchanger has been evaluated in model cells that coexpress the exchanger together with the (protein) Ca indicator aequorin targeted to the different subcellular compartments. Exchanger isoform 1 (NCX1) has been used for experiments in which the effect of its overexpression has been evaluated on the Ca homeostasis in the cytoplasm at large, in the endoplasmic reticulum, in mitochondria, and in the layer of cytoplasm underneath the plasma membrane. The experiments have shown that, despite its very low calcium affinity, the exchanger was active in cells at rest in which calcium is around 100 nM. They have also shown that expressed NCX1 significantly reduced the cytosolic and mitochondrial calcium transients produced by the emptying of the ER calcium stores with InsP3-linked agonists. The effect of the expressed NCX1 on the calcium homeostasis in the domain underneath the plasma membrane was more complex: The exchanger apparently catalyzed a reverse operation, leading to increased penetration of calcium.
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PMID:A study of the activity of the plasma membrane Na/Ca exchanger in the cellular environment. 1250 84

The three Na+/Ca2+ exchanger isoforms, NCX1, NCX2, and NCX3, contain a large cytoplasmic loop that is responsible for the regulation of activity. We have used 347 residues of the loop of NCX2 as the bait in a yeast two-hybrid approach to identify proteins that could interact with the exchanger and regulate its activity. Screening of a human brain cDNA library identified the epsilon and zeta isoforms of the 14-3-3 protein family as interacting partners of the exchanger. The interaction was confirmed by immunoprecipitation and in vitro binding experiments. The effect of the interaction on the homeostasis of Ca2+ was investigated by co-expressing NCX2 and 14-3-3epsilon in HeLa cells together with the recombinant Ca2+ probe aequorin; the ability of cells expressing both NCX2 and 14-3-3epsilon to dispose of a Ca2+ transient induced by an InsP3-producing agonist was substantially decreased, indicating a reduction of NCX2 activity. The 14-3-3epsilon protein also inhibited the NCX1 and NCX3 isoforms. In vitro binding experiments revealed that all three NCX isoforms interacted with multiple 14-3-3 isoforms. 14-3-3 was bound by both phosphorylated and nonphosphorylated NCX, but the phosphorylated form had much higher binding affinity.
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PMID:Inhibitory interaction of the plasma membrane Na+/Ca2+ exchangers with the 14-3-3 proteins. 1667 22