Expression and characterization of protein kinase C-delta.
A cDNA encoding protein kinase C-delta (PKC-delta) 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-delta pseudosubstrate site. The Mr of PKC-delta as determined by SDS/PAGE and immunoblot analysis using anti-(PKC-delta C-terminal) antibodies was 77,000. The enzyme was purified to near homogeneity and showed total dependency on phospholipid and diacylglycerol (or phorbol esters) for activity. Like PKC-epsilon, PKC-delta displays no Ca2+ dependence for activation. The substrate specificity of PCK-delta is similar to that of PKC-epsilon but quite different from other PKCs.
Protein Phosphorylation Laboratory, Imperial Cancer Research Fund, London, England.
A R
AR
Olivier
P J
PJ
Parker
D000595:Amino Acid Sequence; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D004075:Diglycerides / Q000494:pharmacology; D015870:Gene Expression; D015151:Immunoblotting; D007527:Isoenzymes / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D010743:Phospholipids / Q000494:pharmacology; D011493:Protein Kinase C / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D051381:Rats; D013379:Substrate Specificity; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
C011439:1,2-diacylglycerol; D004075:Diglycerides; D007527:Isoenzymes; D010743:Phospholipids; D004247:DNA; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium