Thrombin rapidly induces protein kinase D phosphorylation, and protein kinase C delta mediates the activation.
Thrombin plays a critical role in hemostasis, thrombosis, and inflammation. However, the responsible intracellular signaling pathways triggered by thrombin are still not well defined. We report here that thrombin rapidly and transiently induces activation of protein kinase D (PKD) in aortic smooth muscle cells. Our data demonstrate that protein kinase C (PKC) inhibitors completely block thrombin-induced PKD activation, suggesting that thrombin induces PKD activation via a PKC-dependent pathway. Furthermore, our results show that thrombin rapidly induces PKC delta phosphorylation and that the PKC delta-specific inhibitor rottlerin blocks thrombin-induced PKD activation, suggesting that PKC delta mediates the thrombin-induced PKD activation. Using dominant negative approaches, we demonstrated that expression of a dominant negative PKC delta inhibits the phosphorylation and activation of PKD induced by thrombin, whereas neither PKC epsilon nor PKC zeta affects thrombin-induced PKD activation. In addition, our results of co-immunoprecipitation assays showed that PKD forms a complex with PKC delta in smooth muscle cells. Taken together, the findings of the present study demonstrate that thrombin induces activation of PKD and reveal a novel role of PKC delta in mediating thrombin-induced PKD activation in vascular smooth muscle cells.
Tan
Mingqi
M
Department of Pathology, University of Tennessee, Knoxville, Tennessee 37996, USA.
Xu
Xuemin
X
Ohba
Motoi
M
Ogawa
Wataru
W
Cui
Mei-Zhen
MZ
D000098:Acetophenones / Q000494:pharmacology; D000818:Animals; D001011:Aorta; D001578:Benzopyrans / Q000494:pharmacology; D002478:Cells, Cultured; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D007700:Kinetics; D008301:Maleimides / Q000494:pharmacology; D009131:Muscle, Smooth, Vascular / Q000201:enzymology*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D051745:Protein Kinase C-delta; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D013917:Thrombin / Q000494:pharmacology*
D000098:Acetophenones; D001578:Benzopyrans; D004791:Enzyme Inhibitors; D007093:Imidazoles; D007211:Indoles; D008301:Maleimides; D011725:Pyridines; C085746:rottlerin; C497260:Prkcd protein, rat; C089212:protein kinase D; D011493:Protein Kinase C; D051745:Protein Kinase C-delta; D013917:Thrombin; C070515:bisindolylmaleimide I; C093642:SB 203580; C064758:Ro 31-8220