title: Activation mechanism for CRAC current and store-operated Ca2+ entry: calcium influx factor and Ca2+-independent phospholipase A2beta-mediated pathway.
abstract: Here we tested the role of calcium influx factor (CIF) and calcium-independent phospholipase A2 (iPLA2) in activation of Ca2+ release-activated Ca2+ (CRAC) channels and store-operated Ca2+ entry in rat basophilic leukemia (RBL-2H3) cells. We demonstrate that 1) endogenous CIF production may be triggered by Ca2+ release (net loss) as well as by simple buffering of free Ca2+ within the stores, 2) a specific 82-kDa variant of iPLA2beta and its corresponding activity are present in membrane fraction of RBL cells, 3) exogenous CIF (extracted from other species) mimics the effects of endogenous CIF and activates iPLA2beta when applied to cell homogenates but not intact cells, 4) activation of ICRAC can be triggered in resting RBL cells by dialysis with exogenous CIF, 5) molecular or functional inhibition of iPLA2beta prevents activation of ICRAC, which could be rescued by cell dialysis with a human recombinant iPLA2beta, 6) dependence of ICRAC on intracellular pH strictly follows pH dependence of iPLA2beta activity, and 7) (S)-BEL, a chiral enantiomer of suicidal substrate specific for iPLA2beta, could be effectively used for pharmacological inhibition of ICRAC and store-operated Ca2+ entry. These findings validate and significantly advance our understanding of the CIF-iPLA2-dependent mechanism of activation of ICRAC and store-operated Ca2+ entry.
authors.affiliation: Boston University School of Medicine, Boston, Massachusetts 02118, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Csutora
authors.forename: Vladislav
authors.initials: V
authors.lastname: Zarayskiy
authors.forename: Krisztina
authors.initials: K
authors.lastname: Peter
authors.forename: Francisco
authors.initials: F
authors.lastname: Monje
authors.forename: Tarik
authors.initials: T
authors.lastname: Smani
authors.forename: Sergey I
authors.initials: SI
authors.lastname: Zakharov
authors.forename: Dmitry
authors.initials: D
authors.lastname: Litvinov
authors.forename: Victoria M
authors.initials: VM
authors.lastname: Bolotina
mesh_terms: D000818:Animals; D001792:Blood Platelets / Q000378:metabolism; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism*; D045744:Cell Line, Tumor; D054513:Group IV Phospholipases A2; D010741:Phospholipases A / Q000378:metabolism*; D054467:Phospholipases A2; D051381:Rats
chemical_list: D015220:Calcium Channels; D002352:Carrier Proteins; D010741:Phospholipases A; D054513:Group IV Phospholipases A2; D054467:Phospholipases A2; D002118:Calcium