Regulation of plasmin activity by annexin II tetramer.
Annexin II tetramer (AIIt) is a major Ca(2+)-binding protein of the endothelial cell surface which has been shown to stimulate the tissue plasminogen activator (t-PA)-dependent conversion of plasminogen to plasmin. In the present report, we have examined the regulation of plasmin activity by AIIt. The incubation of plasmin with AIIt resulted in a 95% loss in plasmin activity. SDS-PAGE analysis established that AIIt stimulated the autoproteolytic digestion of plasmin heavy and light chains. The kinetics of AIIt-stimulated plasmin autoproteolysis were first-order, suggesting that binding of plasmin to AIIt resulted in the spontaneous autoproteolysis of the bound plasmin. AIIt did not affect the activity of other serine proteases such as t-PA or urokinase-type plasminogen activator. Furthermore, other annexins such as annexin I, II, V, or VI did not stimulate plasmin autoproteolysis. Increasing the concentration of AIIt on the surface of human 293 epithelial cells increased cell-mediated plasmin autoproteolysis. Thus, in addition to stimulating the formation of plasmin, AIIt also promotes plasmin inactivation. These results therefore suggest that AIIt may function to provide the cell surface with a transient pulse of plasmin activity.
Fitzpatrick
S L
SL
Cancer Biology Research Group, Departments of Biochemistry & Molecular Biology and Oncology, University of Calgary, Calgary, Alberta, Canada T2N 4N1.
Kassam
G
G
Choi
K S
KS
Kang
H M
HM
Fogg
D K
DK
Waisman
D M
DM
D000818:Animals; D017306:Annexin A2 / Q000737:chemistry / Q000502:physiology*; D002417:Cattle; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000737:chemistry; D005341:Fibrinolysin / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D006801:Humans; D006868:Hydrolysis; D007668:Kidney / Q000166:cytology; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism; D013379:Substrate Specificity; D010959:Tissue Plasminogen Activator / Q000378:metabolism
D017306:Annexin A2; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D010958:Plasminogen; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin