PMID: 2494944Apr 1, 1989Paper

Stimulation by fibrinogen of the amidolytic activity of single-chain tissue plasminogen activator

Archives of Biochemistry and Biophysics
T UranoF J Castellino

Abstract

The amidolytic steady-state kinetic properties of a series of recombinant tissue plasminogen activators (rt-PA) have been examined in the presence and absence of the positive effectors fibrinogen (Fg) and native soluble (des-A)-fibrin (sFn). Two-chain (tc) native rt-PA displayed a Km value of 0.50 mM and a kcat value of 13.2 s-1 toward the substrate, H-D-Ile-Pro-Arg-p-nitroanilide (S2288) at 37 degrees. When these same assays were conducted in the presence of Fg or sFn, the Km and kcat values remained essentially the same. On the other hand, the activity of single-chain (sc) rt-PA was significantly increased in the presence of Fg or sFn, by approximately 3.4- to 4-fold, due to alterations in both the Km and kcat of the reaction. Similar results were obtained with rt-PA deletion variants, obtained by site-directed mutagenesis. With rt-PA domain-deletion derivatives, consisting of kringles 1 (K1)-2 (K2)-protease (P), and K2-P, the amidolytic activities of the scrt-PA preparations were significantly stimulated (2.0- to 2.5-fold) by Fg and sFn, a property not shared by the corresponding tcrt-PA. On the other hand, neither the single- nor two-chain derivatives of a deletion mutant containing only the finger (F)-growth factor (E)-P d...Continue Reading

References

Jan 15, 1988·Biochemical and Biophysical Research Communications·T UranoF J Castellino
Jan 1, 1988·Haemostasis·F J CastellinoB A Chibber
Dec 1, 1988·Archives of Biochemistry and Biophysics·J T Radek, F J Castellino
Jul 1, 1972·Archives of Biochemistry and Biophysics·W J Brockway, F J Castellino
Sep 15, 1982·Thrombosis Research·M RånbyP Wallén

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Citations

Dec 11, 2007·Ultrasonics·Azita SoltaniSara Vaezy
May 21, 2018·Journal of Thrombosis and Haemostasis : JTH·T UranoY Suzuki
Jul 17, 2019·Current Drug Targets·Tetsumei UranoFrancis J Castellino
Aug 4, 1995·The Journal of Biological Chemistry·K Tachias, E L Madison
Mar 1, 1991·Archives of Biochemistry and Biophysics·S S Husain

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