Abstract
A 60-kDa human calreticulin was isolated from liver homogenates. The protein was identified as calreticulin by its NH2-terminal amino acid sequence, by its mobility in SDS-PAGE, by its immunoreactivity with anti-calreticulin antibodies, by its Ca2+ binding, and by its localization to isolated ER membranes. In this study we show that Ca2+ binding to calreticulin results in Ca(2+)-dependent aggregation and precipitation of the protein. We also show that calreticulin and calsequestrin bind Zn2+ in 65Zn2+ overlay. In addition we have discovered that calreticulin exhibits a Zn(2+)-dependent interaction with hydrophobic matrix of phenyl-Sepharose that can be utilized in the purification of the protein.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Calcium-Binding Proteins / isolation & purification
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Calcium-Binding Proteins / metabolism*
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Calreticulin
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Chromatography, Affinity
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Chromatography, DEAE-Cellulose
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Dogs
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Electrophoresis, Polyacrylamide Gel
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Endoplasmic Reticulum / metabolism*
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Humans
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Liver / metabolism*
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Molecular Weight
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Muscles / metabolism
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Myocardium / metabolism
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Protein Binding
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Rats
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Ribonucleoproteins / isolation & purification
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Ribonucleoproteins / metabolism*
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Sarcoplasmic Reticulum / metabolism
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Sepharose / analogs & derivatives
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Zinc / metabolism*
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Zinc / pharmacology
Substances
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Calcium-Binding Proteins
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Calreticulin
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Ribonucleoproteins
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phenyl-sepharose
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Sepharose
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Zinc