en
Scientific article
English

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

Published inCell calcium, vol. 14, no. 6, p. 485-492
Publication date1993
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
  • Animals
  • Biological Markers
  • Calcium Channels
  • Calcium-Binding Proteins/ isolation & purification
  • Calreticulin
  • Cell Compartmentation
  • Cell Fractionation
  • Cell Membrane/chemistry
  • Centrifugation, Density Gradient
  • Electrophoresis, Polyacrylamide Gel
  • Endoplasmic Reticulum/chemistry
  • Female
  • Immunoblotting
  • Inositol 1,4,5-Trisphosphate/metabolism
  • Inositol 1,4,5-Trisphosphate Receptors
  • Liver/ chemistry/ultrastructure
  • Microsomes, Liver/chemistry
  • Molecular Weight
  • Rats
  • Rats, Wistar
  • Receptors, Cell Surface/ isolation & purification
  • Receptors, Cytoplasmic and Nuclear
  • Ribonucleoproteins/ isolation & purification
  • Subcellular Fractions/ chemistry
Citation (ISO format)
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. doi: 10.1016/0143-4160(93)90007-s
Identifiers
ISSN of the journal0143-4160
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