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

P B Moore·N Kraus-Friedmann
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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.

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AnimalsCalciumCalcium-Binding ProteinsCalcium-Transporting ATPasesCalmodulinChromatography, AffinityDose-Response Relationship, DrugElectrophoresis, Polyacrylamide GelIn Vitro TechniquesMaleMicrosomes, LiverRatsRats, Inbred StrainsTrifluoperazineJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, Non-P.H.S.Research Support, U.S. Gov't, P.H.S.

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P B Moore, N Kraus-Friedmann. 1983-07-15. Hepatic microsomal Ca2+-dependent ATPase. Calmodulin-dependence and partial purification.. https://doi.org/10.1042/bj2140069

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