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Isolation and characterization of granulocyte lysosomal proteins and study of their effects on the clotting system
Author(s) -
Herion John C.,
Bucher John R.,
Penniall Ralph,
Walker Richard I.,
Baker Mary,
Roberts Harold R.
Publication year - 1979
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/ajh.2830070309
Subject(s) - chemistry , fibrinogen , prekallikrein , kallikrein , biochemistry , trypsin , protease , sephadex , fibrin , high molecular weight kininogen , clotting time , thrombin , kininogen , chromatography , coagulation , enzyme , biology , platelet , immunology , medicine
Lysosomes (granules) of rabbit PMN leukocytes were extracted with either HCl or H 2 SO 4 , and the extracts were chromatographed over Sephadex to separate protein constituents. Some of the low molecular weight cationic proteins homogeneous on SDS PAGE (8% and 12.5% gels) were characterized by electrophoretic mobility in acid gels and by amino acid analysis. A 3,700 dalton polypeptide, rich in arginine and cysteine, prolonged the partial thromboplastin time of normal plasma. In low concentration, this protein shortened the clotting time of pure fibrinogen by thrombin. In high concentration this lysosomal cationic protein precipitated fibrinogen from solution; no fibrinopeptides were released to suggest cleavage of fibrinogen. Fibrinolytic protease activity was detected in crude H 2 SO 4 extracts but not in crude HCl extracts. Two separate plasminogen activators, differing from kallikrein or prekallikrein, were isolated from the H 2 SO 4 lysosomal extract and were partially characterized; neither exhibited proteolytic activity on fibrinogen free of plasminogen.