Localization of the binding site of tissue-type plasminogen activator to fibrin.

Functionally active A and B chains were separated from a two-chain form of recombinant tissue-type plasminogen activator after mild reduction and alkylation. The A chain was found to be responsible for the binding to lysine-Sepharose or fibrin and the B chain contained the catalytic activity of tissue-type plasminogen activator. An extensive reduction of two-chain tissue-type plasminogen activator, however, destroyed both the binding and catalytic activities. A thermolytic fragment, Fr. 1, of tissue-type plasminogen activator that contained a growth factor and two kringle segments retained its lysine binding activity. Additional thermolytic cleavages in the kringle-2 segment of Fr. 1 caused a total loss of the binding activity. These results indicated that the binding site of tissue-type plasminogen activator to fibrin was located in the kringle-2 segment.

[1]  T. Suyama,et al.  Proteolytic cleavage of single-chain pro-urokinase induces conformational change which follows activation of the zymogen and reduction of its high affinity for fibrin. , 1985, The Journal of biological chemistry.

[2]  K Fujikawa,et al.  Amino acid sequence of the heavy chain of human alpha-factor XIIa (activated Hageman factor). , 1985, The Journal of biological chemistry.

[3]  K. Fujikawa,et al.  Proteolytic activation of tissue plasminogen activator by plasma and tissue enzymes , 1984, FEBS letters.

[4]  F. Elgh,et al.  The structure of the human tissue-type plasminogen activator gene: correlation of intron and exon structures to functional and structural domains. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[5]  H. Jörnvall,et al.  Tissue plasminogen activator: peptide analyses confirm an indirectly derived amino acid sequence, identify the active site serine residue, establish glycosylation sites, and localize variant differences. , 1984, Biochemistry.

[6]  J. Zeuthen,et al.  Inactive proenzyme to tissue‐type plasminogen activator from human melanoma cells, identified after affinity purification with a monoclonal antibody. , 1984, The EMBO journal.

[7]  L. Holmberg,et al.  Immunological characterisation of plasminogen activators in the human vessel wall. , 1983, Journal of clinical pathology.

[8]  J. Griffin,et al.  Isolation and functional characterization of the active light chain of activated human blood coagulation factor XI. , 1983, The Journal of biological chemistry.

[9]  D. Collen,et al.  Measurement of human tissue-type plasminogen activator by a two-site immunoradiometric assay. , 1983, The Journal of laboratory and clinical medicine.

[10]  P. Seeburg,et al.  Cloning and expression of human tissue-type plasminogen activator cDNA in E. coli , 1983, Nature.

[11]  J. Griffin,et al.  Isolation and functional properties of the heavy and light chains of human plasma kallikrein. , 1982, The Journal of biological chemistry.

[12]  M. Rånby,et al.  Purification and identification of two structural variants of porcine tissue plasminogen activator by affinity adsorption on fibrin. , 1982, Biochimica et biophysica acta.

[13]  L. Flohé,et al.  The primary structure of high molecular mass urokinase from human urine. The complete amino acid sequence of the A chain. , 1982, Hoppe-Seyler's Zeitschrift fur physiologische Chemie.

[14]  D. Collen,et al.  Purification and characterization of the plasminogen activator secreted by human melanoma cells in culture. , 1981, The Journal of biological chemistry.

[15]  D. Rijken,et al.  Relationship between tissue plasminogen activator and the activators in blood and vascular wall. , 1980, Thrombosis research.

[16]  J. Conard,et al.  Modifications of α2-Plasmin Inhibitor During Treatments by a Defibrinating Agent , 1979, Thrombosis and Haemostasis.

[17]  B. Wiman,et al.  The specific interaction between plasminogen and fibrin. A physiological role of the lysine binding site in plasminogen. , 1977, Thrombosis research.

[18]  T. Astrup,et al.  Differences in the Binding to Fibrin of Urokinase and Tissue Plasminogen Activator , 1972, Thrombosis and Haemostasis.

[19]  D. Deutsch,et al.  Plasminogen: Purification from Human Plasma by Affinity Chromatography , 1970, Science.

[20]  I. Goldfine,et al.  Radioimmunoassay of human plasminogen and plasmin. , 1969, The Journal of laboratory and clinical medicine.

[21]  J. M. Minor,et al.  Simultaneous selectivity optimization of mobile and stationary phases in reversed-phased liquid chromatography for isocratic separations of phenylthiohydantoin amino acid derivatives , 1985 .

[22]  R. Allen,et al.  Isolation and Properties of Human Vascular Plasminogen Activator , 1977, Thrombosis and Haemostasis.