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Title

Organization of Ca2+ stores in myeloid cells: association of SERCA2b and the type-1 inositol-1,4,5-trisphosphate receptor

Authors
Favre, C. J.
Jerstrom, P.
Stendhal, O.
Huggler, E.
Published in Biochemical Journal. 1996, vol. 316 ( Pt 1), p. 137-142
Abstract In this study, we have analysed the relationship between Ca2+ pumps and Ins(1,4,5)P3-sensitive Ca2+ channels in myeloid cells. To study whether sarcoplasmic/endoplasmic reticulum Ca(2+)-ATPase (SERCA)-type Ca(2+)-ATPases are responsible for Ca2+ uptake into Ins(1,4,5)P3-sensitive Ca2+ stores, we used the three structurally unrelated inhibitors thapsigargin, 2,5-di-t-butylhydroquinone and cyclopiazonic acid. In HL-60 cells, all three compounds precluded formation of the phosphorylated intermediate of SERCA-type Ca(2+)-ATPases. They also decreased, in parallel, ATP-dependent Ca2+ accumulation and the amount of Ins(1,4,5)P3-releasable Ca2+. Immunoblotting with subtype-directed antibodies demonstrated that HL-60 cells contain the Ca2+ pump SERCA2 (subtype b), and the Ca(2+)-release-channel type-1 Ins(1,4,5)P3 receptor. In subcellular fractionation studies, SERCA2 and type-1 Ins(1,4,5)P3 receptor co-purified. Immunofluorescence studies demonstrated that both type-1 Ins(1,4,5)P3 receptor and SERCA2 were evenly distributed throughout the cell in moving neutrophils. During phagocytosis both proteins translocated to the periphagosomal space. Taken together, our results suggest that in myeloid cells (i) SERCA-type Ca(2+)-ATPases function as Ca2+ pumps of Ins(1,4,5)P3-sensitive Ca2+ stores, and (ii) SERCA2 and type-1 Ins(1,4,5)P3 receptor reside either in the same or two tightly associated subcellular compartments.
Keywords Calcium/ metabolismCalcium Channels/ metabolismCalcium-Transporting ATPases/antagonists & inhibitors/ metabolismEndoplasmic Reticulum/ enzymologyEnzyme Inhibitors/pharmacologyHL-60 CellsHumansHydroquinones/pharmacologyIndoles/pharmacologyInositol 1,4,5-Trisphosphate ReceptorsKineticsNeutrophils/ enzymologyPhagocytosisPhosphorylationReceptors, Cytoplasmic and Nuclear/ metabolismSarcoplasmic Reticulum/ enzymologyTerpenes/pharmacologyThapsigarginVanadates/pharmacology
Identifiers
PMID: 8645196
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Other version: http://www.biochemj.org/bj/316/0137/3160137.pdf
Structures
Research group Obésité et syndrome métabolique (803)
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(ISO format)
FAVRE, C. J. et al. Organization of Ca2+ stores in myeloid cells: association of SERCA2b and the type-1 inositol-1,4,5-trisphosphate receptor. In: Biochemical Journal, 1996, vol. 316 ( Pt 1), p. 137-142. https://archive-ouverte.unige.ch/unige:7201

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Deposited on : 2010-06-21

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