Protein kinase B is regulated in platelets by the collagen receptor glycoprotein VI.
Phosphoinositide 3-kinase (PI3K) is a critical component of the signaling pathways that control the activation of platelets. Here we have examined the regulation of protein kinase B (PKB), a downstream effector of PI3K, by the platelet collagen receptor glycoprotein (GP) VI and thrombin receptors. Stimulation of platelets with collagen or convulxin (a selective GPVI agonist) resulted in PI3K-dependent, and aggregation independent, Ser(473) and Thr(308) phosphorylation of PKBalpha, which results in PKB activation. This was accompanied by translocation of PKB to cell membranes. The phosphoinositide-dependent kinase PDK1 is known to phosphorylate PKBalpha on Thr(308), although the identity of the kinase responsible for Ser(473) phosphorylation is less clear. One candidate that has been implicated as being responsible for Ser(473) phosphorylation, either directly or indirectly, is the integrin-linked kinase (ILK). In this study we have examined the interactions of PKB, PDK1, and ILK in resting and stimulated platelets. We demonstrate that in platelets PKB is physically associated with PDK1 and ILK. Furthermore, the association of PDK1 and ILK increases upon platelet stimulation. It would therefore appear that formation of a tertiary complex between PDK1, ILK, and PKB may be necessary for phosphorylation of PKB. These observations indicate that PKB participates in cell signaling downstream of the platelet collagen receptor GPVI. The role of PKB in collagen- and thrombin-stimulated platelets remains to be determined.
Barry
Fiona A
FA
School of Animal & Microbial Sciences, University of Reading, Whiteknights, Reading RG6 6AJ, United Kingdom.
Gibbins
Jonathan M
JM
D064413:3-Phosphoinositide-Dependent Protein Kinases; D001792:Blood Platelets / Q000201:enzymology*; D003094:Collagen / Q000502:physiology; D003435:Crotalid Venoms / Q000378:metabolism; D006801:Humans; D016023:Integrins / Q000502:physiology*; D037181:Lectins, C-Type; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt; D038981:Receptors, Collagen; D000099200:Scaffold Protein ILK
D064413:3-Phosphoinositide-Dependent Protein Kinases; D003094:Collagen; D003435:Crotalid Venoms; D016023:Integrins; D037181:Lectins, C-Type; D017346:Protein Serine-Threonine Kinases; D011518:Proto-Oncogene Proteins; D051057:Proto-Oncogene Proteins c-akt; D038981:Receptors, Collagen; D000099200:Scaffold Protein ILK; C028890:convulxin; C578562:PDPK1 protein, human