Plasminogen interaction with platelets: the importance of carboxyterminal lysines.
BACKGROUND
Thrombin stimulation enhances plasminogen binding to platelets and promotes platelet-dependent plasmin generation. The objective of this study was to determine whether carboxyterminal lysines (C-lysines) are important for these processes, as they are in other cell types.


METHODS
125I-plasminogen and varying concentrations of unlabeled plasminogen were added to washed platelets that were either resting or stimulated with thrombin, thrombin receptor activating peptide, or ADP. In some experiments the platelets were digested with carboxypeptidase B to remove C-lysines. Platelet-dependent plasmin generation was also studied by adding plasminogen and tissue plasminogen activator to platelet suspensions and monitoring the conversion of a plasmin specific chromogenic substrate. The cells were either resting or stimulated with thrombin, thrombin receptor activating peptide, or ADP. The effect of the thrombin inhibitor lepirudin and the plasmin inhibitor aprotinin on plasminogen binding and the appearance of C-lysines was also investigated.


RESULTS
Thrombin, but not thrombin receptor activating peptide or ADP, stimulated high-affinity binding of plasminogen and greatly promoted platelet-dependent plasmin generation. Digestion with carboxypeptidase B eliminated thrombin-induced high-affinity binding and reduced thrombin-induced plasmin generation by increasing the Michaelis constant. Lepirudin, but not aprotinin, inhibited thrombin-stimulated plasminogen binding to platelets.


CONCLUSIONS
C-terminal lysines are necessary for high-affinity binding of plasminogen to platelets and for platelet-supported plasmin generation. The origin of the C-lysines is not clear, but they may result from a direct effect of thrombin, rather than an intermediate enzyme such as plasmin.
Horne
McDonald K
MK
Hematology Service, Department of Laboratory Medicine, W. G. Magnuson Clinical Center, National Institutes of Health, Rm 2C306, Building 10, NIH, Bethesda, MD 20892, USA. mhorne@mail.cc.nih.gov
Merryman
Paula K
PK
Cullinane
Ann M
AM
D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D043424:Carboxypeptidase B / Q000494:pharmacology; D005341:Fibrinolysin / Q000096:biosynthesis; D006801:Humans; D007457:Iodine Radioisotopes; D007700:Kinetics; D008239:Lysine / Q000737:chemistry / Q000378:metabolism*; D010958:Plasminogen / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology
D007457:Iodine Radioisotopes; D010958:Plasminogen; D043424:Carboxypeptidase B; D013917:Thrombin; D005341:Fibrinolysin; D008239:Lysine