Feedback regulation of endothelial cell surface plasmin generation by PKC-dependent phosphorylation of annexin A2.
In response to blood vessel injury, hemostasis is initiated by platelet activation, advanced by thrombin generation, and tempered by fibrinolysis. The primary fibrinolytic protease, plasmin, can be activated either on a fibrin-containing thrombus or on cells. Annexin A2 (A2) heterotetramer (A2·p11)(2) is a key profibrinolytic complex that assembles plasminogen and tissue plasminogen activator and promotes plasmin generation. We now report that, in endothelial cells, plasmin specifically induces activation of conventional PKC, which phosphorylates serine 11 and serine 25 of A2, triggering dissociation of the (A2·p11)(2) tetramer. The resulting free p11 undergoes ubiquitin-mediated proteasomal degradation, thus preventing further translocation of A2 to the cell surface. In vivo, pretreatment of A2(+/+) but not A2(-/-) mice with a conventional PKC inhibitor significantly reduced thrombosis in a carotid artery injury model. These results indicate that augmentation of fibrinolytic vascular surveillance by blockade of serine phosphorylation is A2-dependent. We also demonstrate that plasmin-induced phosphorylation of A2 requires both cleavage of A2 and activation of Toll-like receptor 4 on the cell surface. We propose that plasmin can limit its own generation by triggering a finely tuned "feedback" mechanism whereby A2 becomes serine-phosphorylated, dissociates from p11, and fails to translocate to the cell surface.
Department of Cell and Developmental Biology, Weill Cornell Medical College, New York, New York 10065, USA.
Kai-Li
KL
He
Guangzhi
G
Sui
Huabao
H
Xiong
M Johan
MJ
Broekman
Bihui
B
Huang
Aaron J
AJ
Marcus
Katherine A
KA
Hajjar
D000818:Animals; D017306:Annexin A2 / Q000378:metabolism*; D042783:Endothelial Cells / Q000378:metabolism*; D025461:Feedback, Physiological; D005341:Fibrinolysin / Q000096:biosynthesis*; D051379:Mice; D018345:Mice, Knockout; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D055503:Protein Multimerization; D021381:Protein Transport; D012694:Serine / Q000378:metabolism
D017306:Annexin A2; D012694:Serine; D011493:Protein Kinase C; D005341:Fibrinolysin