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Title: Conversion of protein kinase C from a Ca/sup 2 +/-dependent to an independent form of phorbol ester-binding protein by digestion with trypsin

Tryptic fragments of protein kinase C containing the kinase (45 KDa) and phorbol ester-binding activity (38 KDa) were separated by Mono O column chromatography. The purified phorbol ester-binding fragment exhibits a higher affinity for phosphatidylserine than the native enzyme but comparable Kd for (/sup 3/H)phorbol 12,13-dibutyrate as the native enzyme. This proteolytic fragment binds phorbol ester equally efficient either in the presence or absence of Ca/sup 2 +/ and the addition of the kinase fragment did not restore the Ca/sup 2 +/-requirement for the binding. These results indicate that protein kinase C is composed of two functionally distinct units which can be expressed independently after limited proteolysis with trypsin.
Authors:
;
Publication Date:
OSTI Identifier:
6937280
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochem. Biophys. Res. Commun.; (United States); Journal Volume: 1
Research Org:
National Institute of Child Health and Human Development, Bethesda, MD
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; PHORBOL ESTERS; CHEMICAL BONDS; PHOSPHOTRANSFERASES; BIOCHEMICAL REACTION KINETICS; CALCIUM; RATS; TRACER TECHNIQUES; TRITIUM COMPOUNDS; TRYPSIN; ALKALINE EARTH METALS; ANIMALS; CARCINOGENS; ELEMENTS; ENZYMES; ESTERS; HYDROLASES; ISOTOPE APPLICATIONS; KINETICS; LABELLED COMPOUNDS; MAMMALS; METALS; ORGANIC COMPOUNDS; PEPTIDE HYDROLASES; PHOSPHORUS-GROUP TRANSFERASES; REACTION KINETICS; RODENTS; SERINE PROTEINASES; TRANSFERASES; VERTEBRATES 550201* -- Biochemistry-- Tracer Techniques