The major objectives of this proposal are to study the basic biological processes involved in the regulation and control of the blood fibrinolytic system in health and disease. They include the biosynthesis of urokinase-type (u-PA) and tissue-type (t-PA) plasminogen activators in normal mammalian tissues, e.g. monkey and baboon kidney, liver and spleen. These studies include the isolation of specific mRNA, the isolation of translation products by specific immunoprecipitation methods, and the characterization of the activator forms. Abnormal, variant, plasminogens will be isolated form the plasma of patients with thrombotic diseases, and possibly from full-term newborn umbilical cord plasmas, and attempts will be made to determine the molecular defect(s). Plasma u-PA and t-PA forms will be isolated by affinity chromatographic methods, from normal subjects, patients with arterial and venous thrombotic diseases, patients with secondary fibrinolytic states (DIC), and cancer patients in remission treated with stanozolol (an anabolic steroid). The plasma u-PA and t-PA forms (zymogen and enzyme) will be characterized by various chemical, physical and immunologic methods. Their role in the mechanism of physiological and pathological fibrinolysis will be investigated. Activator binding domains, and fibrin-binding domains, on both normal and abnormal plasminogens will be identified using peptide mapping methods by HPLC, and specific antibody probes. Recombinant hybrid activator enzymes will be prepared from both the plasminogen fibrin-binding domains(A chains) and the u-PA/t-PA active center domains (B chains), and their properties will be compared to the parent u-PA and t-PA enzymes.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL034276-03
Application #
3347007
Study Section
Hematology Subcommittee 2 (HEM)
Project Start
1984-06-01
Project End
1988-05-31
Budget Start
1986-06-01
Budget End
1987-05-31
Support Year
3
Fiscal Year
1986
Total Cost
Indirect Cost
Name
Michael Reese Hosp & Medical Center (Chicago)
Department
Type
DUNS #
City
Chicago
State
IL
Country
United States
Zip Code
60616
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Robbins, K C; Boreisha, I G; Godwin, J E (1991) Abnormal plasminogen Maywood I. Thromb Haemost 66:575-80
Robbins, K C (1990) Classification of abnormal plasminogens: dysplasminogenemias. Semin Thromb Hemost 16:217-20
Lee, P P; Wohl, R C; Boreisha, I G et al. (1988) Kinetic analysis of covalent hybrid plasminogen activators: effect of CNBr-degraded fibrinogen on kinetic parameters of Glu1-plasminogen activation. Biochemistry 27:7506-13
Robbins, K C; Barlow, G H; Nguyen, G et al. (1987) Comparison of plasminogen activators. Semin Thromb Hemost 13:131-8
Robbins, K C; Boreisha, I G (1987) A covalent molecular weight approximately 92,000 hybrid plasminogen activator derived from human plasmin fibrin-binding and tissue plasminogen activator catalytic domains. Biochemistry 26:4661-7
Summaria, L; Boreisha, I; Barlow, G H et al. (1987) The isolation and characterization of a ternary human plasmin B-chain-streptokinase-plasminogen complex. Thromb Haemost 58:772-7
Scharrer, I M; Wohl, R C; Hach, V et al. (1986) Investigation of a congenital abnormal plasminogen, Frankfurt I, and its relationship to thrombosis. Thromb Haemost 55:396-401
Toki, N; Sumi, H; Sasaki, K et al. (1985) Transport of urokinase across the intestinal tract of normal human subjects with stimulation of synthesis and/or release of urokinase-type proteins. J Clin Invest 75:1212-22