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Title

Inositol 1,4,5-trisphosphate binding sites copurify with the putative Ca-storage protein calreticulin in rat liver

Authors
Enyedi, P.
Szabadkai, G.
Spat, A.
Published in Cell Calcium. 1993, vol. 14, no. 6, p. 485-492
Abstract Rat liver was homogenized and subjected to differential centrifugation. When the low speed nuclear pellet was processed on a Percoll gradient, plasma membrane markers and Ins(1,4,5)P3 binding activity purified together. The high speed (microsomal) fraction was subfractionated by sucrose density gradient centrifugation, resulting in 10-fold enrichment of [32P]-Ins(1,4,5)P3 binding. In the sucrose density gradient fractions there was an inverse relationship between the enrichment of plasma membrane markers and Ins(1,4,5)P3 binding sites. Endoplasmic reticulum markers showed a moderate enrichment in the fractions displaying high Ins(1,4,5)P3 binding activity. Calcium binding proteins in the homogenate and in the microsomal subfractions were separated by SDS/PAGE. A 60 kD protein, stained metachromatically with Stains-All was identified as calreticulin with immunoblotting. Its enrichment pattern was similar to that of Ins(1,4,5)P3 binding sites, indicating the co-existence of these two elements of Ca(2+)-metabolism in the same intracellular compartment in the liver.
Keywords AnimalsBiological MarkersCalcium ChannelsCalcium-Binding Proteins/ isolation & purificationCalreticulinCell CompartmentationCell FractionationCell Membrane/chemistryCentrifugation, Density GradientElectrophoresis, Polyacrylamide GelEndoplasmic Reticulum/chemistryFemaleImmunoblottingInositol 1,4,5-Trisphosphate/metabolismInositol 1,4,5-Trisphosphate ReceptorsLiver/ chemistry/ultrastructureMicrosomes, Liver/chemistryMolecular WeightRatsRats, WistarReceptors, Cell Surface/ isolation & purificationReceptors, Cytoplasmic and NuclearRibonucleoproteins/ isolation & purificationSubcellular Fractions/ chemistry
Identifiers
PMID: 8395340
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ENYEDI, P. et al. Inositol 1,4,5-trisphosphate binding sites copurify with the putative Ca-storage protein calreticulin in rat liver. In: Cell Calcium, 1993, vol. 14, n° 6, p. 485-492. https://archive-ouverte.unige.ch/unige:9317

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Deposited on : 2010-07-12

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