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Title

Purification of an inositol 1,4,5-trisphosphate-binding calreticulin-containing intracellular compartment of HL-60 cells

Authors
Favre, C.
Spat, A.
Cerny, E.
Published in Biochemical Journal. 1992, vol. 281 ( Pt 3), p. 651-656
Abstract To investigate the identity of Ins(1,4,5)P3-sensitive intracellular Ca2+ stores in myeloid cells, we have developed a method that yields subcellular fractions highly enriched in Ins(1,4,5)P3 binding. HL-60 cells were disrupted by nitrogen cavitation, and subcellular fractions were obtained by differential centrifugation, followed by Percoll- and sucrose-density-gradient separations. A subcellular fraction enriched 26-fold in Ins(1,4,5)P3-binding sites was obtained. This fraction showed no enrichment in plasma-membrane markers and only a comparatively moderate enrichment (7-fold) in endoplasmic-reticulum markers. The ratio between specific enrichment of Ins(1,4,5)P3 binding and endoplasmic-reticulum markers in the different fractions varied over 50-fold, from less than 0.1 to greater than 5. The purified Ins(1,4,5)P3-binding fraction was enriched to a similar extent (27-fold) in the putative intravesicular Ca(2+)-storage protein calreticulin. Our results favour the concept of a distinct Ins(1,4,5)P3-binding, calreticulin-containing compartment (i.e. the calciosome) in HL-60 cells.
Keywords Calcium-Binding Proteins/ analysisCalreticulinCell FractionationCell Membrane/metabolismCentrifugation, Density GradientEndoplasmic Reticulum/chemistry/ metabolismHumansImmunoblottingInositol 1,4,5-Trisphosphate/ metabolism/pharmacologyNADPH-Ferrihemoprotein Reductase/analysisSulfatases/analysisTumor Cells, Cultured
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PMID: 1536644
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Other version: http://www.biochemj.org/bj/281/0651/2810651.pdf
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VAN DELDEN, Christian et al. Purification of an inositol 1,4,5-trisphosphate-binding calreticulin-containing intracellular compartment of HL-60 cells. In: Biochemical Journal, 1992, vol. 281 ( Pt 3), p. 651-656. https://archive-ouverte.unige.ch/unige:7626

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Deposited on : 2010-06-21

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