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J. Biochem, 1988, Vol. 104, No. 4 643-647
© 1988 Japanese Biochemical Society

research-article

Synthesis of Recombinant Human Single-Chain Urokinase-Type Plasminogen Activator Variants Resistant to Plasmin and Thrombin

Toshio Miyake1, Yasuo Hibino, You-ichi Kobayashi, Midori Yokoyama, Ken Watabe and Muneki Ohmori

Sagami Chemical Research Center Sagamihara, Kanagawa 229

Single-chain urokinase-type plasminogen activator (scu-PA), a potential therapeutic reagent for thrombosis, is activated in plasma by plasmin. The activated enzyme is further digested by plasmin to generate low-molecular-weight urokinase (LMW-UK), which has no affinity for fibrin. To circumvent this dual effect of plasmin, we synthesized in Escherichia coli a variant of scu-PA, which is not converted to LMW-UK on treatment with plasmin. In another variant, the activation cleavage site was modified such that activation by plasmin was slowed down and that inactivation by thrombin was greatly diminished. The combination of these variants may be applicable as an effective thrombolytic reagent for clinical use.

1Present address: Biotechnology Research Laboratory. TOSOH Corporation, Ayase, Kanagawa 252.


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