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Thromb Haemost 1987; 57(01): 082-086
DOI: 10.1055/s-0038-1651067
Original Articles
Schattauer GmbH Stuttgart

Mapping of Epitopes on Human Tissue-Type Plasminogen Activator with Recombinant Deletion Mutant Proteins

Anton-Jan van Zonneveld
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Depts. of Molecular Biology and Autoimmune Diseases, Amsterdam, The Netherlands
,
Harry Veerman
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Depts. of Molecular Biology and Autoimmune Diseases, Amsterdam, The Netherlands
,
Just P J Brakenhoff
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Depts. of Molecular Biology and Autoimmune Diseases, Amsterdam, The Netherlands
,
Lucien A Aarden
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Depts. of Molecular Biology and Autoimmune Diseases, Amsterdam, The Netherlands
,
Jean-Francois Cajot
*   The Swiss Institute for Experimental Cancer Research, Epalinges, Switzerland
,
Hans Pannekoek
The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Depts. of Molecular Biology and Autoimmune Diseases, Amsterdam, The Netherlands
› Author Affiliations
Further Information

Publication History

Received 22 August 1986

Accepted after revision 25 November 1986

Publication Date:
06 July 2018 (online)

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Summary

An antigen assay based on a monoclonal antibody directed against the light chain of tissue-type plasminogen activator (t-PA) was developed to quantify seven recombinant (r) t-PA deletion mutant proteins. These recombinant proteins were then employed to map different epitopes on t-PA which interact with a panel of twenty-three monoclonal anti-t-PA antibodies. Twenty were directed against domains on the heavy chain, two against the “finger” domain, three against the “epidermal growth factor-like” domain, five against the kringle 1 domain, and ten against the kringle 2 domain. Only three monoclonal anti-t-PA antibodies interact with the light chain. The finding that the epitopes of each of the monoclonals could be determined with the deletion mutant proteins supports the hypothesis of autonomous folding of structural domains and emphasizes the validity of the use of the recombinant t-PA-deletion mutant proteins for structure-function studies.

Keywords

Recombinant tissue-type plasminogen activator - Monoclonal antibodies - Epitopes - Domain structure
 

  • References

  • 1 Binder BR, Spragg J, Austen KF. Purification and characterization of human vascular plasminogen activator derived from blood vessel perfusates. J Biol Chem 1979; 254: 1998-2003
    PubMedSearch in Google Scholar
  • 2 Wallen P, Riinby M, Bergsdorf N, Kok P. Purification and characterization of tissue plasminogen activator; on the occurrence of two different forms and their enzymatic properties. In: Progress in Fibrinolysis. Davidson JF, Nilsson IM, Asted B. (eds.), vol. 5 pp. 229-232 Edinburgh: Churchill Livingstone; 1981
    Search in Google Scholar
  • 3 Pennica D, Holmes WE, Kohr WJ, Harkins RN, Vehar GA, Ward CA, Bennet WF, Yelverton E, Seeburg PH, Heyneker HL, Goeddel DV. Cloning and expression of human tissue-type plasminogen activator cDNA in E. coli . Nature 1983; 301: 214-222
    CrossrefPubMedSearch in Google Scholar
  • 4 Banyai L, Varadi A, Patthy L. Common evolutionary origin of fibrinbinding structures of fibronectin and tissue-type plasminogen activator. FEBS Lett 1983; 163: 37-41
    CrossrefPubMedSearch in Google Scholar
  • 5 Verstraete H, Bory M, Collen D, Erbel R, Lennane RJ, Mathey D, Michels HR, Schartl M, Uebis R, Bernard R, Brower RW, De Bono DP, Huhmann W, Lubsen J, Meyer J, Rutsch W, Schmidt W, von Essen R. Randomnised trial of intravenous recombinant tissue-type plasminogen activator versus intravenous streptokinase in acute myocardial infarction. The Lancet 1985; 1: 842-847
    PubMedSearch in Google Scholar
  • 6 Williams DO, Borer J, Braunwald E, Chesebro JH, Cohen LS, Dalen J, Dodge HT, Francis CK, Knatterud G, Ludbrook P, Markis JE, Mueller H, Desvigne-Nickens P, Passamani ER, Powers ER, Rao AK, Roberts R, Ross A, Ryan TJ, Sobel BE, Winniford M, Zaret B. Intravenous recombinant tissue-type plasminogen activator in patients with acute myocardial infarction: a report from the NHLBI thrombolysis in myocardial infarction trial. Circulation 1986; 73: 338-346
    CrossrefPubMedSearch in Google Scholar
  • 7 Thorsen S, Glas-Greenwalt P, Astrup T. Differences in binding to fibrin of urokinase and tissue-type plasminogen activator. Thromb Diath Haemorrh 1972; 28: 65-74
    PubMedSearch in Google Scholar
  • 8 Hoylaerts M, Rijken DC, Lijnen HR, Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator. J Biol Chem 1982; 257: 338-346
    PubMedSearch in Google Scholar
  • 9 MacDonald ME, Van Zonneveld A-J, Pannekoek H. Functional analysis of the human tissue-type plasminogen activator protein: the light chain. Gene 1986; 42: 59-67
    CrossrefPubMedSearch in Google Scholar
  • 10 Van Zonneveld A-J, Veerman H, Pannekoek H. Autonomous functions of structural domains of human tissue-type plasminogen activator. Proc Natl Acad Sci USA 1986; 83: 4670-4674
    CrossrefPubMedSearch in Google Scholar
  • 11 Kagitani H, Tagawa M, Hatanaka K, Ikari T, Saito A, Bando H, Okada K, Matsuo O. Expression in E. coli of finger-domain lacking tissue-type plasminogen activator with high fibrin affinity. FEBS Lett 1985; 189: 145-149
    CrossrefPubMedSearch in Google Scholar
  • 12 Mattler L, Marks CA, Bang NU. Factors influencing tissue-type plasminogen activator binding to fibrin and fibrin-dependent plasminogen activation. Thrombos Haemostas 1985; 54: 266 (Abstr)
    PubMedSearch in Google Scholar
  • 13 MacGregor IR, Micklem LR, James K, Pepper DS. Characterization of epitopes on human tissue plasminogen activator recognized by a group of monoclonal antibodies. Thromb Haemostas 1985; 53: 45-50
    PubMedSearch in Google Scholar
  • 14 Wallen P, Bergsdorf N, Pohl G, Stigbrand T. Differentiation between one-chain and two-chain tissue type plasminogen activator by monoclonal antibodies. Haemostasis 1984; 14: 42
    PubMedSearch in Google Scholar
  • 15 Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 1975; 256: 495-497
    CrossrefPubMedSearch in Google Scholar
  • 16 Aarden L, Lansdorp P, De Groot E. A growth factor for hybridoma cells produced by human monocytes. Lymphokines 1985; 10: 175-185
    PubMedSearch in Google Scholar
  • 17 Lopata MA, Cleveland DW, Sollner-Webb B. High level transient expression of a chloroamphenicol acetyl transferase gene by DEAE mediated DNA transfection coupled with a dimethyl sulfoxide or glycerol shock treatment. Nucleic Acids Res 1984; 12: 5707-5717
    CrossrefPubMedSearch in Google Scholar
  • 18 Hunter WM, Greenwood FC. Preparation of 131lodine-labeled human growth hormone of high specific activity. Nature 1962; 194: 495-496
    CrossrefPubMedSearch in Google Scholar
  • 19 Patthy L. Evolution of the proteases of blood coagulation and fibrinolysis by assembly from modules. Cell 1985; 41: 657-663
    CrossrefPubMedSearch in Google Scholar
  • 20 Gilbert W. Genes-in-pieces revisited. Science 1985; 228: 823-824
    CrossrefPubMedSearch in Google Scholar