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

Autor: Elzbieta Huggler, Daniel Pablo Lew, Cécile J. Favre, Petra Jerström, Karl-Heinz Krause, Michelangelo Foti, Olle Stendhal
Rok vydání: 1996
Předmět:
Calcium/ metabolism
Thapsigargin
SERCA
Calcium-Transporting ATPases/antagonists & inhibitors/ metabolism
Indoles
Neutrophils
Receptors
Cytoplasmic and Nuclear
HL-60 Cells
Calcium-Transporting ATPases
Endoplasmic Reticulum
Biochemistry
chemistry.chemical_compound
Phagocytosis
Humans
Inositol 1
4
5-Trisphosphate Receptors
Inositol
Enzyme Inhibitors
Phosphorylation
Receptor
Receptors
Cytoplasmic and Nuclear/ metabolism
Molecular Biology
ddc:616
Enzyme Inhibitors/pharmacology
Hydroquinones/pharmacology
Terpenes/pharmacology
Voltage-dependent calcium channel
Indoles/pharmacology
Sarcoplasmic Reticulum/ enzymology
Vanadates/pharmacology
Terpenes
Endoplasmic reticulum
Cell Biology
Neutrophils/ enzymology
Molecular biology
Hydroquinones
Kinetics
Sarcoplasmic Reticulum
chemistry
Immunology
Calcium Channels/ metabolism
Calcium
Calcium Channels
Cell fractionation
Vanadates
Cyclopiazonic acid
Endoplasmic Reticulum/ enzymology
Research Article
Zdroj: Biochemical Journal, Vol. 316 ( Pt 1) (1996) pp. 137-142
ISSN: 0264-6021
Popis: 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 Ca2+-ATPase (SERCA)-type Ca2+-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 Ca2+-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 Ca2+-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 Ca2+-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.
Databáze: OpenAIRE
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