The 2.3 Å Crystal Structure of the Catalytic Domain of Recombinant Two-chain Human Tissue-type Plasminogen Activator

Journal of Molecular Biology

Volumn 258, Issue 1, 1996, Pages 117-135

  Lamba, Doriano ^a   Bauer, Margit ^a   Huber, Robert ^a   Fischer, Stephan ^b   Rudolph, Rainer ^b   Kohnert, Ulrich ^b   Bode, Wolfram ^a  

^a MAX PLANCK INSTITUTE OF BIOCHEMISTRY   (Germany)

^b F HOFFMANN LA ROCHE LTD   (Switzerland)

Author keywords

Catalytic domain; Crystal structure; Fibrinolysis; Serine proteinase; Tissue type plasminogen activator

Indexed keywords

DISULFIDE; RECOMBINANT PROTEIN; SERINE PROTEINASE; TISSUE PLASMINOGEN ACTIVATOR; TRYPSIN;

ARTICLE; CRYSTAL STRUCTURE; ENZYME ACTIVE SITE; ENZYME CONFORMATION; HUMAN; KRINGLE DOMAIN; MOLECULAR MODEL; PRIORITY JOURNAL; PROTEIN DOMAIN; X RAY CRYSTALLOGRAPHY; AMINO ACID SEQUENCE; BINDING SITE; CATALYSIS; CHEMISTRY; GENE DELETION; GENETICS; MOLECULAR GENETICS; PROTEIN TERTIARY STRUCTURE; SEQUENCE ALIGNMENT;

AMINO ACID SEQUENCE; BINDING SITES; CATALYSIS; CRYSTALLOGRAPHY, X-RAY; HUMANS; MOLECULAR SEQUENCE DATA; PROTEIN STRUCTURE, TERTIARY; RECOMBINANT PROTEINS; SEQUENCE ALIGNMENT; SEQUENCE DELETION; TISSUE PLASMINOGEN ACTIVATOR;

EID: 0029665065     PISSN: 00222836     EISSN: None     Source Type: Journal    
DOI: 10.1006/jmbi.1996.0238     Document Type: Article

References (104)

1. Andreasen, P. A., Petersen, L. C. & Dano, K. (1991). Diversity in catalytic properties of single chain and two chain tissue-type plasminogen activator. Fibrinolysis, 5, 207-215.
2. Bachmann, F. (1994). Molecular aspects of plasminogen, plasminogen activators and plasmin. In Haemostasis and Thrombosis (Bloom, A. L., Forbes, C. D., Thomas, D. P., Tuddenham, E. G. D., eds), vol. 1, pp. 575-613, Churchill, Livingstone, Edinburgh, London, Madrid, Melbourne, New York and Tokyo.
3. Bakker, A. H. F., Rehberg, E. F., Marotti, K. R. & Verheijen, J. H. (1995). The position of the structurally autonomous kringle 2 domain influences the functional features of tissue-type plasminogen. Protein Eng. 8, 293-300.
4. Bennett, W. F., Paoni, N. F., Keyt, B. A., Botstein, D. Jones, A. J. S., Presta, L., Wurm, F. M. & Zoller, M. J. (1991). High resolution analysis of functional determinants on human tissue-type plasminogen activator. J. Biol. Chem. 266, 5191-5201.
5. Bergum, P. W. & Gardell, S. J. (1992). Vampire bat salivary plasminogen activator exhibits a strict and fastidious requirement for polymeric fibrin as its cofactor, unlike human tissue-type plasminogen activator. J. Biol. Chem. 267, 17726-17731.
6. Bernstein, F. C., Koetzle, T. F., Williams, G. J. B., Meyer, E. F., Jr, Brice, M. D., Rodgers, J. R., Kennard, O., Shimanouchi, T, & Tasumi, M. (1977). The Protein Data Bank: a computer-based archival file for macromolecular structures. J. Mol. Biol. 11, 535-542.
7. Blundell, T. L., Sibanda, B. L., Sternberg, M. J. E. & Thorton, J. M. (1987). Knowledge-based prediction of protein structures and the design of novel molecules. Nature, 326, 347-352.
8. Bode, W. (1979). The transition of bovine trypsinogen to a trypsin-like state upon strong ligand binding. II. The binding of the pancreatic trypsin inhibitor and of isoleucine-valine and of sequentially related peptides to trypsinogen and to p-guanidinobenzoate-trypsinogen. J. Mol. Biol. 127, 357-374.
9. Bode, W. & Huber, R. (1986). Crystal structures of pancreatic serine endopeptidases. In Molecular and Cellular Basis of Digestion (Desnuelle P., ed.), pp. 213-234, Elsevier, Amsterdam, New York, Oxford.
10. Bode, W. & Huber, R. (1992). Natural proteinase inhibitors and their interaction with proteinases. Eur. J. Biochem. 204, 433-451.
11. Bode, W. & Schwager, P. (1975). The refined crystal structure of bovine β-trypsin at 1.8 Å resolution. II. Crystallographic refinement, calcium binding site, benzamidine binding site and active site at pH 7. J. Mol. Biol. 98, 693-717.
12. Bode, W., Fehlhammer, H. & Huber, R. (1976). Crystal structure of bovine trypsinogen at 1.8 Å resolution. I. Data collection, application of Patterson search techniques and preliminary structural interpretation. J. Mol. Biol. 106, 325-335.
13. Bode, W., Schwager, P. & Huber, R. (1978). The transition of bovine trypsinogen to a trypsin-like state upon strong ligand binding. J. Mol. Biol. 118, 99-112.
14. Bode, W., Mayr, I., Baumann, U., Hube, R., Stone, S. R. & Hofsteenge, J. (1989). The refined 1.9 Å crystal structure of human α-thrombin: interaction with D-Phe-Pro-Arg chloromethylketone and significance of the Tyr-Pro-Pro-Trp insertion segment. EMBO J. 8, 3467-3475.
15. Bode, W., Turk, D. & Karshikov, A. (1992). The refined 1.9 Å X-ray crystal structure of D-Phe-Pro-Arg chloromethylketone-inhibited human α-thrombin: structure analysis, overall structure, electrostatic properties, detailed active-site geometry and structure-function relationships. Protein Sci. 1, 426-471.
16. Bolognesi, M., Gatti, G., Menegatti, E., Guarneri, M., Marquart, M., Papamokos, E. & Huber, R. (1982). Three-dimensional structure of the complex between pancreatic secretory trypsin inhibitor (Kazal-type) and trypsinogen at 1.8 Å resolution. Structure solution, crystallographic refinement and preliminary interpretation. J. Mol. Biol. 162, 839-868.
17. Brandstetter, H., Bauer, M., Huber, R., Lollar, P. & Bode, W. (1995). X-ray structure of clotting factor IXa: active site and module structure related to Xase activity and haemophilia B. Proc. Natl Acad. Sci. USA, 92, 9797-9800.
18. Bringmann, P., Gruber, D., Liese, A., Toschi, L., Krätzschmar, J., Schleuning, W.-D. & Donner, P. (1995). Structural features mediating fibrin selectivity of vampire bat plasminogen activators. J. Biol. Chem. 270, 25596-25603.
19. Brünger, A. T. (1992a). X-PLOR, verion 3.1. A system for X-ray crystallography and NMR. Yale University Press, New Haven.
20. Brünger, A. T. (1992b). Free R value: a novel statistic quantity for assessing the accuracy of crystal structures. Nature, 355, 472-474.
21. Byeon, I.-J. L. & Llinas, M. (1991). Solution structure of the tissue-type plasminogen activator kringle 2 domain complexed to 6-aminohexanoic acid and antifibrinolytic drug. J. Mol. Biol. 222, 1035-1051.
22. 1H-NMR. Biochemistry, 34, 2739-2750.
23. CCP4 Suite (1994). Collaborative Computational Project, Number 4. Acta Crystallog. sect. D, 50, 760-763.
24. Chmielewska, J., Rånby, M. & Wiman, B. (1988). Kinetics of the inhibition of plasminogen activators by the plasminogen-activator inhibitor. Biochem. J. 251, 327-332.
25. Collen, D. & Lijnen, H. R. (1995). Molecular basis of fibrinolysis, as relevant for thrombolytic therapy Thromb. Haemostasis, 74, 167-171.
26. Cool, D. E. & MacGillivray, R. T. A. (1987). Characterization of the human blood coagulation factor XII gene. J. Biol. Chem. 262, 13662-13673.
27. De Vos, A. M., Ultsch, M. H., Kelley, R. F., Padmanabhan, K., Tulinsky, A., Westbrook, M. L. & Kossiakoff, A. A. (1992). Crystal structure of the kringle 2 domain of tissue plasminogen activator at 2.4 Å resolution. Biochemistry, 31, 270-279.
28. Ding, L., Coombs, G. S., Strandberg, L., Navre, M., Corey D. R. & Madison, E. L. (1995). Origins of the specificity of tissue-type plasminogen activator. Proc. Natl Acad Sci. USA, 92, 7627-7631.
29. Dodd, I., Browne, M. J., Chapman, C. G., Cronk, D., Hastwell, C., Wilson, S. & Robinson, J. H. (1993). Deletion of residues K296-G302 from the slowly-cleared tissue-type plasminogen activator t-PA del(G) leads to partial loss of plasminogen activating activity Biologicals, 21, 269-274.
30. 1H nuclear magnetic resonance. J. Mol. Biol. 225, 821-833.
31. Eastman, D., Wurm, F. M., Van Reis, R. & Higgins, D. L. (1992). A region of tissue plasminogen activator that affects plasminogen activation differentially with various fibrin(ogen)-related stimulators. Biochemistry, 31, 419-422.
32. Engh, R. A. & Huber, R. (1991). Accurate bond and angle parameters for X-ray protein structure refinement. Acta Crystallog. sect. A, 47, 392-400.
33. Fehlhammer, H., Bode, W. & Huber, R. (1977). Crystal structure of bovine trypsinogen at 1.8 A resolution. II. Crystallographic refinement, refined crystal structure and comparison with bovine trypsin. J. Mol. Biol. 111, 415-438.
34. Freer, S. T., Kraut, J., Robertus, J. D., Wright, H. T. & Xuong, N. H. (1970). Chymotrypsinogen: 2.5-Å crystal structure, comparison with α-chymotrypsin, and implications for zymogen activation. Biochemistry, 9, 1997-2009.
35. Gardell, S. J., Duong, L. T., Diehel, R. E., York, J. D., Hare, T. R., Register, R. B., Jacobs, J. W., Dixon, R. A. F. & Friedman, P. A. (1989). Isolation, characterization, and cDNA cloning of a vampire bat salivary plasminogen activator. J. Biol. Chem. 264, 17947-17952.
36. Gething, M.-J., Adler, B., Boose, J.-A., Gerard, R. D., Madison, E. L., McGookey, D., Meidell, R. S., Roman, L. M. & Sambrook, J. (1988). Variants of human tissue-type plasminogen activator that lack specific structural domains of the heavy chain. EMBO J. 7, 2731-2740.
37. Goldberger, G., Bruns, G. A. P., Rits, M., Edge, M. D. & Kwiatkowski, D. J. (1987). Human complement factor I: analysis of cDNA-derived primary structure and assignment of its gene to chromosome 4. J. Biol. Chem. 262, 10065-10071.
38. Grailhe, P., Nieuwenhuizen, W. & Angles-Cano, E. (1994). Study of tissue-type plasminogen activator binding sites on fibrin using distinct fragments of fibrinogen. Eur. J. Biochem. 219, 961-967.
39. Hartley, B. S. & Shotton, D. M. (1971). Pancreatic elastase. In The Enzymes (Boyer P. D., ed.), pp. 323-373, Academic Press, New York and London.
40. Heckel, A. & Hasselbach, K. M. (1988). Prediction of the three-dimensional structure of the enzymatic domain of t-PA. J. Comput-Aid Mol. Des. 2, 7-14.
41. Hedstrom, L., Szilagy, L. & Rutter, W. J. (1992). Converting trypsin to chymotrypsin: the role of surface loops. Science, 255, 1249-1253.
42. Heussen, C., Joubert, F. & Dowdle, E. B. (1984). Purification of human tissue plasminogen activator with Erythrina trypsin inhibitor. J. Biol. Chem. 259, 11635-11638.
43. Higgins, D. L. & Vehar, G. A. (1987). Interaction of one-chain and two-chain tissue plasminogen activator with intact and plasmin-degraded fibrin. Biochemistry, 26, 7786-7791.
44. Higgins, D. L., Young, S. L., Powers, D. B. & Anderson, S. (1992). A zymogenic tissue plasminogen-activator variant - the Phe305His mutation suppresses fibrin(ogen) stimulated plasminogen activation by one chain t-PA. Fibrinolysis, 6, 161-166.
45. Horrevoets, A. J. G., Tans, G., Smilde, A. E., van Zonneveld, A. J. & Pannekoek, H. (1993). Thrombin-variable region 1 (VR1). J. Biol. Chem. 268, 779-782.
46. Hoylaerts, M., Rijken, D. C., Lijnen, H. R. & Collen, D. (1982). Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin. J. Biol. Chem. 257, 2912-2919.
47. Hu, C.-K., Kohnert, U., Wilhelm, O., Fischer, S. & Llinas, M. (1994). Tissue-type plasminogen activator domain-deletion mutant BM 06.022: modular stability inhibitor binding, and activation cleavage. Biochemistry, 3, 11760-11766.
48. Huber, R. & Bode, W. (1978). Structural basis of the activation and action of trypsin. Acc. Chem. Res. 11, 114-122.
49. Huber, R., Kukla, D., Bode, W., Schwager, P., Bartels, K., Deisenhofer, J. & Steigemann, W. (1974). Structure of the complex formed by bovine trypsin and bovine pancreatic trypsin inhibitor. II. Crystallographic refinement at 1.9 Å resolution. J. Mol. Biol. 89, 73-101.
50. Jones, T. A., Zou, J.-Y., Cowan, S. W & Kjeldgaard, M. (1991). Improved methods for building protein models in eletron density maps and location of errors in these models. Acta Crystallog. sect. A, 47, 110-119.
51. Joubert, F.J. & Dowdle, E.B. (1987). The primary structure of the inhibitor of tissue plasminogen activator found in the seeds of Erythrina caffra. Thromb. Haemostas, 57, 356-360.
52. Kohnert, U., Rudolph, R., Verheijen, J. H., Weening-Verhoeff, E. J. D., Stern, A., Opitz, U. Martin, U., Lill, H., Prinz, H., Lechner, M., Kresse, G.-B. Buckel, P. & Fischer, S. (1992). Biochemical properties of the kringle 2 and protease domains are maintained in the refolded t-PA deletion variant BM 06.022. Protein Eng. 5, 93-100.
53. Kohnert, U., Horsch, B. & Fischer, S. (1993). A variant of tissue plasminogen activator (t-PA) comprised of the kringle 2 and the protease domain shows a significant difference in the in vitro rate of plasmin formation as compared to the recombinant human t-PA from transformed Chinese hamster ovary cells. Fibrinolysis, 7, 365-372.
54. Kohnert, U., Hellerbrand, K., Martin, U., Stern, A., Popp, F. & Fischer, S. (1996). The recombinant Escherichia coli-derived protease domain of tissue-type plasminogen activator is a potent and fibrin specific fibrinolytic agent. Fibrinolysis, 10, in the press.
55. Kraulis, P. J. (1991). MOLSCRIPT: a program to produce both detailed and schematic plots of protein structures. J. Appl. Crystallog. 24, 946-950.
56. Laskowski, R. A., MacArthur, M. W., Moss, D. S. & Thorton, J. M. (1993). PROCHECK: a program to check the stereochemical quality of protein structures. J. Appl. Crystallog. 26, 283-291.
57. Leslie, A. G. W. (1994). Mosflm User Guide, Mosflm version 5.20. MRC Laboratory of Molecular Biology, Cambridge, UK.
58. 2Cl, a synthetic urokinase inhibitor, with single-chain and two-chain forms of urokinase-type plasminogen activator. Eur. J. Biochem. 162, 351-356.
59. Lijnen, H. R., Bachmann, F., Collen, D., Ellis, V., Pannekoek, H., Rijken, D. C. & Thorsen, S. (1994). Mechanisms of plasminogen activation. J. Intern. Med. 236, 415-424.
60. Luzzati, V. (1952). Traitement statistique des erreurs dans la determination des structures cristallines. Acta Crystallog. 5, 802-810.
61. Madison, E. L. (1994). Probing structure-function relationships of tissue-type plasminogen activator by site-specific mutagenesis. Fibrinolysis, 8, 221-236.
62. Madison, E. L., Goldsmith, E. J., Gerard, R. D., Gething, M. J. H. & Sambrook, J. F. (1989). Serpin-resistant mutants of human tissue-type plasminogen activator. Nature, 339, 721-724.
63. Madison, E. L., Goldsmith, E. J., Gerard, R. D., Gething, M.-J. H., Sambrook, J. F. & Bassel-Duby, R. S. (1990a). Amino acid residues that affect interaction of tissue-type plasminogen activator with plasminogen activator inhibitor 1. Proc. Natl Acad. Sci. USA, 87, 3530-3533.
64. Madison, E. L., Goldsmith, E. J., Gething, M.-J., Sambrook, J. F. & Gerard, R. D. (1990b). Restoration of serine protease-inhibitor interaction by protein engineering. J. Biol. Chem. 265, 21423-21426.
65. Madison, E. L., Kobe, A., Gething, M.-J., Sambrook, J. F. & Goldsmith, E. J. (1993). Converting tissue plasminogen activator to a zymogen: a regulatory triad of Asp-His-Ser. Science, 262, 419-421.
66. Madison, E. L., Coombs, G. S. & Corey, D. R. (1995). Substrate specificity of tissue-type plasminogen activator. J. Biol. Chem. 270, 7558-7562.
67. Mangel, W. F., Lin, B. & Ramakrishnan, V. (1990). Characterization of an extremely large ligand-induced conformational change in plasminogen. Science, 48, 69-73.
68. Margossian, S. S., Slayter, H. S., Kaczmarek, E. & McDonagh, J. (1993). Physical characterization of recombinant tissue plasminogen activator. Biochim. Biophys. Acta, 1163, 250-256.
69. Merrit, E. A. & Murphy, M. E. P. (1994). Raster3D version 2.0 - A program for photorealistic molecular graphics. Acta Chrysallog. sect. D, 50, 869-873.
70. Miyazawa, K., Shimomura, T., Kitamura, A., Kondo, J., Morimoto, Y. & Kitamura, N. (1993). Molecular cloning and sequence analysis of the cDNA for a human serine protease responsible for activation of hepatocyte growth factor. J. Biol. Chem. 268, 10024-10028.
71. 1 alteplase, and streptokinase following intravenous bolus injection in a rabbit model of carotid artery thrombosis. Fibrinolysis, 7, 284-290.
72. Navaza, J. (1994). AMoRe: an automated package for molecular replacement. Acta Crystallog. sect. A, 50, 157-163.
73. Nicholls, A., Bharadwaj, R. & Honig, B. (1993). GRASP-graphical representation and analysis of surface properties. Biophys. J. 64, A166.
74. Nienaber, V. L., Young, S. L., Birktof, J. J., Higgins, D. L. & Berliner, L. J. (1992). Conformational similarities between one-chain and two-chain tissue plasminogen activator (t-PA): implications to the activation mechanism on one-chain t-PA. Biochemistry, 31, 3852-3861.
75. Novokhatny, V. V., Ingham, K. C. & Medved, L. V. (1991). Domain structure and domain-domain interactions of recombinant tissue plasminogen activator. J. Biol. Chem. 266, 12944-13002.
76. Onesti, S., Brick, P. & Blow, D. M. (1991). Crystal structure of a Kunitz-type trypsin inhibitor from Erythrina caffra seeds. J. Mol. Biol. 217, 153-176.
77. Padmanabhan, K., Padmanabhan, K. P., Tulinsky, A., Park, C. H., Bode, W., Huber, R., Blankenship, D. T., Cardin, A. D. & Kisiel, W. (1993). Structure of human des(1-45) factor Xa at 2.2 Å resolution. J. Mol. Biol. 232, 947-966.
78. Padmanabhan, K., Wu, T. P., Ravichandran, K. G. & Tulinsky, A. (1994). Kringle-kringle interactions in multimer kringle structures. Protein Sci. 3, 898-910.
79. Pannekoek, H., Lijnen, H. R. & Loskutoff, D. J. (1990). Recommendations for the nomenclature of mutant fibrinolytic genes and their proteins. Thromb. Haemostas, 64, 600-603.
80. Paoni, N. F., Chow, A. M., Peña, L. C., Keyt, B. A., Zoller, M. J. & Bennett, W. F. (1993a). Making tissue-type plasminogen activator more fibrin specific. Protein Eng. 6, 529-534.
81. Paoni, N. F., Keyt, B. A., Refino, C. J., Chow, A. M., Nguyen, H. V., Bereleau, L. T., Badillo, J., Peña, L. C., Brady, K., Wurm, F. M., Ogez, J. & Bennett, W. F. (1993b). A slow clearing, fibrin-specific, PAI-1 resistant variant of t-PA (T103N, KHRR 296-299 AAAA). Thromb. Haemostas, 70, 307-312.
82. Pennica, D., Holmes, W. E., Kohr, W. J., Harkins, R. N., Vehar, G. A., Ward, C. A., Bennett, W. F., Yelverton, E., Seeburg, P. H., Heyneker, H. L., Goeddel, D. V. & Collen, D. (1983). Cloning and expression of human tissue-type plasminogen activator cDNA in E. coli. Nature, 301, 214-221.
83. Perona, J. J. & Craik, C. S. (1995). Structural basis of substrate specificity in the serine proteases. Protein Sci. 4, 337-360.
84. Petersen, L. C., Boel, E., Johannessen, O. & Foster, D. (1990). Quenching of the amidolytic activity of one-chain tissue-type plasminogen activator by mutation of lysine-416. Biochemistry, 23, 3451-3457.
85. Pohl, G., Källström, M., Bergsdorf, N., Wallen, P. & J. Jörnvall, H. (1984). 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. Biochemistry, 23, 3701-3707.
86. Read, J. M. (1986). Improved Fourier coefficients for maps using phases from partial structures with errors. Acta Crystallog. sect. A, 42, 140-149.
87. Reilly, C. F. & Hutzelmann, J. E. (1992). Plasminogen activator inhibitor-1 binds to fibrin and inhibits tissue-type plasminogen activator-mediated fibrin dissolution. J. Biol. Chem. 267, 17128-17135.
88. Schechter, I. & Berger, A. (1967). On the size of the active site in proteases. I. Papain. Biochem. Biophys. Res. Commun. 27, 157-162.
89. Shohet, R. V., Spitzer, S., Madison, E. L., Bassel-Duby, R., Gething, M.-J. & Sambrook, J. F. (1994). Inhibitor-resistant tissue-type plasminogen activator: an improved thrombolytic agent in vitro. Thromb. Haemostas, 71, 124-128.
90. Sippl, M. J. (1993). Recognition of errors in the three-dimensional structures of proteins. Proteins: Struct. Funct. Genet, 17, 355-362.
91. Smith, B. O., Downing, A. K., Driscoll, P. C., Dudgeon, T. J. & Campbell, I. D. (1995). The solution structure and backbone dynamics of the fibronectin type I and epidermal growth factor-like pair of modules of tissue-type plasminogen activator. Structure, 3, 823-833.
92. Spraggon, G., Phillips, C., Nowak, U. K., Ponting, C. P., Saunders, D., Dobson, C. M., Stuart, D. I. & Jones, E. Y. (1995). The crystal structure of the catalytic domain of human urokinase-type plasminogen activator. Structure, 3, 681-691.
93. Steigemann, W. (1991). Recent advances in the PROTEIN program system for the X-ray structure analysis of biological macromolecules. In Crystallographic Computing (Moras, D., Podjarny, A. D. & Thiery J. C., eds), vol. 5, pp. 115-125, Oxford University Press, Oxford.
94. Straßburger, W., Wollmer, A., Pitts, J. E., Glover, I. D., Tickle, I. J., Blundell, T. L., Steffens, G. J., Günzler, W. A., Ötting, F. & Flohe, L. (1983). Adaptation of plasminogen activator sequences to known protease structures. FEBS Letters, 157, 219-223.
95. Strandberg, L. & Madison, E. L. (1995). Variants of tissue-type plasminogen activator with substantially enhanced response and selectivity toward fibrin co-factors. J. Biol. Chem. 270, 23444-23449.
96. Stürzebecher, J., Neumann, U., Kohnert, U., Kresse, G.-B. & Fischer, S. (1992). Mapping of the catalytic site of CHO-t-PA and the t-PA variant BM 06.022 by synthetic inhibitors and substrates. Protein Sci. 1, 1007-1013.
97. Tachias, K. & Madison, E. L. (1995). Variants of tissue-type plasminogen activator which display substantially enhanced stimulation by fibrin. J. Biol. Cell, 270, 18319-18322.
98. Teixera, A. V., Dowdle, E. B. D. & Botes, D. P. (1994). Site-directed mutagenesis of the synthetic Erythrina trypsin/tissue plasminogen activator (t-PA) inhibitor encoding-gene to compare the interaction of Erythrina and soybean trypsin inhibitor with t-PA. Biochim. Biophys. Acta, 1217, 23-28.
99. Turk, D. (1992). PhD thesis, Technische Universität München.
100. Van Zonneveld, A. J., Veerman, H. & Pannekoek, H. (1986). Autonomous functions of structural domains of human tissue-type plasminogen activator. Proc. Natl Acad. Sci. USA, 83, 4670-4674.
101. Verheijen, J. H., Caspers, M. P. M., Chang, G. T. G., De Munk, G. A. W., Pouwels, P. H. & Enger-Valk, B. E. (1986). Involvement of finger domain and kringle 2 domain of tissue-type plasminogen activator in fibrin binding and stimulation of activity by fibrin. EMBO J. 5, 3525-3530.
102. Wallen, P., Pohl, G., Bersdorf, N., Rånby M., Ny, T. & Jörnvall, H. (1983). Purification and characterization of a melanoma cell plasminogen activator. Eur. J. Biochem. 182, 681-686.
103. Wang, D., Bode, W. & Huber, R. (1985). Bovine chymotrypsinogen A. X-ray crystal structure analysis and refinement of a new crystal form at 1.8 Å resolution. Mol. Biol. 185, 595-624.
104. Wittwer, A. J., Howard, S. C., Carr, L. S., Harakas, N. K., Feder, J., Parekh, R. B. Rudd, P. M., Dwek, R. A. & Rademacher, T. W. (1989). Effects of N-glycosylation on in vitro activity of Bowes melanoma and human colon fibroblast derived tissue plasminogen activator. Biochemistry, 28, 7662-7669.