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

P. Enyedi, G. Szabadkai, K. H. Krause, D. P. Lew, A. Spät

Research output: Contribution to journalArticle

22 Citations (Scopus)

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 Ca2+-metabolism in the same intracellular compartment in the liver.

Original languageEnglish
Pages (from-to)485-492
Number of pages8
JournalCell Calcium
Volume14
Issue number6
DOIs
Publication statusPublished - 1993

Fingerprint

Calreticulin
Inositol 1,4,5-Trisphosphate
Sucrose
Binding Sites
Cell Membrane
Calcium-Binding Proteins
Density Gradient Centrifugation
Liver
Centrifugation
Immunoblotting
Endoplasmic Reticulum
Polyacrylamide Gel Electrophoresis
Proteins
Percoll
stains-all

ASJC Scopus subject areas

  • Cell Biology
  • Endocrinology

Cite this

Inositol 1,4,5-trisphosphate binding sites copurify with the putative Ca-storage protein calreticulin in rat liver. / Enyedi, P.; Szabadkai, G.; Krause, K. H.; Lew, D. P.; Spät, A.

In: Cell Calcium, Vol. 14, No. 6, 1993, p. 485-492.

Research output: Contribution to journalArticle

@article{0274b4fe4e2640de9495b63c14d315d9,
title = "Inositol 1,4,5-trisphosphate binding sites copurify with the putative Ca-storage protein calreticulin in rat liver",
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 Ca2+-metabolism in the same intracellular compartment in the liver.",
author = "P. Enyedi and G. Szabadkai and Krause, {K. H.} and Lew, {D. P.} and A. Sp{\"a}t",
year = "1993",
doi = "10.1016/0143-4160(93)90007-S",
language = "English",
volume = "14",
pages = "485--492",
journal = "Cell Calcium",
issn = "0143-4160",
publisher = "Churchill Livingstone",
number = "6",

}

TY - JOUR

T1 - Inositol 1,4,5-trisphosphate binding sites copurify with the putative Ca-storage protein calreticulin in rat liver

AU - Enyedi, P.

AU - Szabadkai, G.

AU - Krause, K. H.

AU - Lew, D. P.

AU - Spät, A.

PY - 1993

Y1 - 1993

N2 - 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 Ca2+-metabolism in the same intracellular compartment in the liver.

AB - 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 Ca2+-metabolism in the same intracellular compartment in the liver.

UR - http://www.scopus.com/inward/record.url?scp=0027184321&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0027184321&partnerID=8YFLogxK

U2 - 10.1016/0143-4160(93)90007-S

DO - 10.1016/0143-4160(93)90007-S

M3 - Article

VL - 14

SP - 485

EP - 492

JO - Cell Calcium

JF - Cell Calcium

SN - 0143-4160

IS - 6

ER -