A recombinant prourokinase chimera was constructed by introduction of Lys-Gly-Asp-Trp-motif between Gly118 and Ile119 among the kringle domain. The structure of designed protein was predicted and simulated. The recombinant prourokinase chimera was produced in insect cell sf9 with baculovirus-expression vector and existed as active form. Chimera protein was purified by affinity chromatography coupled with antibody. The special activity of the chimera was 90,000 IU/mg detected by fibrin plate determination. It was also shown that chimera inhibited ADP-induced platelet aggregation in a concentration depenent manner. These results showed the prourokinase chimera exhibited not only high fibrinolytic activity but also had anti-thrombosis function.
 Gunzler, V.A., Steffens, G.J., Otting, F., et al. (1982) Structural relationship between human high and low molecular mass urokinase. Hoppe-Seyler’s Zeitschrift für physiologische Chemie, 363, 133-141. doi:10.1515/bchm2.1982.363.1.133
 Husain, S.S., Gurewich, V. and Liinsik, B. (1983) Purification and partial characterization of a single-chain high-molecular-weight form of urokinase. Archives of Biochemistry and Biophysics, 220, 31-38. doi:10.1016/0003-9861(83)90383-1
 Homes, W.E., Pennica, D., Blaber, M., Rey, M.W., Guenzler, W.M., Staffen, G.J. and Heyneker, H.L. (1985) Cloning and expression of the gene for prourokinase in Escherichia coli. Biotechnology, 3, 923-929. doi:10.1038/nbt1085-923
 Nelles, L., Lijnen, H.R., Collen, D. and Holmes, W.E., (1987) Characterization of recombinant human single chain urokinase-type plasminogen activator mutants. Journal of Biological Chemistry, 262, 5682-5689.
 Scarborough, R., Rose, J.W., Hsu, M.A., Phillips, D.R., Fried, V.A., Campbell, A.M., Nannizzi, L. and Charo, I.F. (1991) Barbourin. A GPIIb/IIIa-specific integrin antagonist from the venom of Sistrurus m barbouri. Journal of Biological Chemistry, 566, 9359-9362.
 Minoux, H., Chipot, C., Brown, D. and Maigret, B. (2000) Structural analysis of the KGD sequence loop of barbourin an alphaIIbbeta3-specific disintegrin. Journal of Computer-Aided Molecular Design, 14, 317-327. doi:10.1023/A:1008182011731
 Plow, E.F., Marguerie, G. and Ginsberg, M. (1987) Fibrinogen, fibrinogen receptors, and the peptides that inhibit these interactions. Biochemical Pharmacology, 36, 4036-4040. doi:10.1016/0006-2952(87)90558-2
 Maeda, S. (1989) Gene transfer vectors of a baculovirus, Bombyx mori unclear polyhedrosis virus, and their use for expression of foregn genes in insect cells [C]. In: Mitsuhashi, J., Ed., Invertebrate Cell System Appliactions, CRC Press, Boca Raton, 163.
 Bradford, M.M. (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye-binding. Analytical Biochemistry, 72, 248-54. doi:10.1016/0003-2697(76)90527-3
 Dennis, M.S., Henzel, W.J., Pitti, R.M., Lipari, M.T., Napier, M.A., Deisher, T., Bunting, S. and Lazarus, R.A. (1989) Platelet glycoprotein IIb-IIIa protein antagonists from snake venoms: Evidence for a family of platelet-aggregation inhibitors. Proceedings of the National Academy of Sciences, 87, 2471-2475. doi:10.1073/pnas.87.7.2471
 Shebuski, R.J., Stabilito, I.J., Sitko, G.R. and Polokoff, M.H. (1990) Acceleration of recombinant tissue-type plasminogen activator induced thrombolysis and prevention of reocclusion by the combination of heparin and the Arg-Gly-Asp-containing peptide bitistatin in a canine model of coronary thrombosis. Circulation, 82, 169-177. doi:10.1161/01.CIR.82.1.169
 Shebuski, R.J., Berry, D.E., Bennett, D.B., Storer, B.L., Ali, F. and Samanen, J. (1989) Demonstration of Ac-ArgGly-Asp-Ser-NH2 as an antiaggregatory agent in the dog intracoronary administration. Thrombos Haemostas, 61, 183-188.