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PMID: 6225427 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Hepatic microsomal Ca2+-dependent ATPase. Calmodulin-dependence and partial purification.

The Biochemical journal ·Vol. 214 ·No. 1 ·1983-07-15 ·Pages 69-75

Moore PB, Kraus-Friedmann N

Abstract

The hepatic microsomal fraction contains tightly bound calmodulin as demonstrated by affinity chromatography. When this calmodulin was partially removed by EGTA treatment (0.5 mM-EGTA), the uptake of 45Ca2+ by the microsomal vesicles was stimulated by added calmodulin and inhibited by trifluoperazine (TFP). The Ca2+-dependent ATPase was partially purified on a calmodulin column. This partial purification resulted in a 500-fold increase in the specific activity of the enzyme when measured in the presence of added calmodulin. Antibodies prepared against calmodulin prevented this stimulatory effect. The fraction eluted from the calmodulin column contained several protein bands indicating that the specific activity of the Ca2+-dependent ATPase is probably still underestimated. There are likely to be other calmodulin-sensitive processes present in the hepatic microsomal fraction.

MeSH Terms
Animals Calcium/metabolism Calcium-Binding Proteins/pharmacology Calcium-Transporting ATPases/isolation & purification,metabolism Calmodulin/pharmacology Chromatography, Affinity Dose-Response Relationship, Drug Electrophoresis, Polyacrylamide Gel In Vitro Techniques Male Microsomes, Liver/drug effects,enzymology Rats Rats, Inbred Strains Trifluoperazine/pharmacology
Chemicals
Calcium-Binding Proteins Calmodulin Trifluoperazine Calcium-Transporting ATPases Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Moore P B
Kraus-Friedmann N
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22 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj2140069
Published
1983-07-15
Pages
69-75
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1152211
Subset
IM
Grants
NIGMS NIH HHS · GM29323 · United States
External Links
PubMed source DOI: 10.1042/bj2140069 PMC Full Text View journal details
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