Molecular cloning and characterization of protein kinase D: a target for diacylglycerol and phorbol esters with a distinctive catalytic domain.
A serine/threonine protein kinase that binds phorbol esters and diacylglycerol (named protein kinase D, PKD) has been identified. PKD contains membrane localization signals and a cysteine-rich repeat sequence homologous to that seen in the regulatory domain of protein kinase C (PKC). A bacterially expressed N-terminal domain of PKD exhibited high-affinity phorbol ester binding activity (Kd = 35 nM). The diacylglycerol analog 1-oleoyl-2-acetylglycerol inhibited phorbol ester binding in a dose-dependent manner. The catalytic domain of PKD contains all characteristic sequence motifs of serine protein kinases but shows only a low degree of sequence similarity to PKCs. The highest identity is with the catalytic domain of myosin light-chain kinase from Dictyostelium (41%). The bacterially expressed catalytic domain of PKD efficiently phosphorylated the exogenous peptide substrate syntide 2 in serine but did not catalyze significant phosphorylation of a variety of other substrates used by PKCs and other major second messenger regulated kinases. PKD may be an unusual component in the transduction of diacylglycerol and phorbol ester signals.
Valverde
A M
AM
Imperial Cancer Research Fund, London, United Kingdom.
Sinnett-Smith
J
J
Van Lint
J
J
Rozengurt
E
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D003001:Cloning, Molecular; D004075:Diglycerides / Q000502:physiology; D015870:Gene Expression; D005796:Genes; D051379:Mice; D008969:Molecular Sequence Data; D010703:Phorbol Esters / Q000494:pharmacology; D011493:Protein Kinase C / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D011955:Receptors, Drug / Q000235:genetics*; D015290:Second Messenger Systems; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D013379:Substrate Specificity
D029742:Caenorhabditis elegans Proteins; D002352:Carrier Proteins; D004075:Diglycerides; D010703:Phorbol Esters; D012333:RNA, Messenger; D011955:Receptors, Drug; C083141:phorbol ester binding protein; C027776:phorbol ester receptor; C089212:protein kinase D; D017346:Protein Serine-Threonine Kinases; D011493:Protein Kinase C