Open AccessExpression and characterization of protein kinase C‐δ
Author(s) -
OLIVIER Andrée R.,
PARKER Peter J.
Publication year - 1991
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1991.tb16248.x
Subject(s) - protein kinase c , diacylglycerol kinase , complementary dna , microbiology and biotechnology , transfection , enzyme , biology , prkcq , cdna library , chemistry , biochemistry , phorbol ester , gene
A cDNA encoding protein kinase C‐δ (PKC‐δ) was isolated from a rat brain library. The coding region was subcloned into the expression vector pmt2 and transfected into COS‐1 cells. Expression of the protein led to an 11‐fold increase in activity as determined with a synthetic peptide based on the PKC‐δ pseudosubstrate site. The M r of PKC‐δ as determined by SDS/PAGE and immunoblot analysis using anti‐(PKC‐δ C‐terminal) antibodies was 77000. The enzyme was purified to near homogeneity and showed total dependency on phospholipid and diacylglycerol (or phorbol esters) for activity. Like PKC‐ɛ, PKC‐δ displays no Ca 2+ dependence for activation. The substrate specificity of PCK‐δ is similar to that of PKC‐ɛ but quite different from other PKCs.