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Activation of plasminogen by single-chain urokinase or by two-chain
urokinase--a demonstration that single-chain urokinase has a low catalytic
activity (pro-urokinase)
R Pannell and V Gurewich
Single-chain urokinase (SC-UK) has an intrinsic amidolytic activity, as
measured with synthetic substrate (Kabi S-2444; pyro-Glu-Gly-Arg-
pNitroanalide), which was found to be 0.1% to 0.2% that of its plasmin-
activated derivative, two-chain UK (TC-UK). A study of the reaction of
SC-UK with plasminogen is complicated by the effect of the reaction
product, plasmin, on both reactants. The resultant generation of TC-UK and
Lys-plasminogen produces secondary reactions which greatly augment
plasminogen activation. To confine enzymatic activity to the primary
reaction, after pretreatment to eliminate trace TC-UK contaminants, SC- UK
was incubated with Glu- or Lys-plasminogen in the presence of aprotinin
(500 KIU/mL) to inhibit generated plasmin and dansyl-glutamyl-
glycyl-arginyl-chloromethylketone (20 mumol/L), which irreversibly
inhibited TC-UK but not SC-UK. Analysis by reducing sodium dodecyl
sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed a
plasminogen-activating activity for SC-UK that was approximately 0.4% that
of TC-UK. Both SC-UK and TC-UK preferentially activated Lys- plasminogen
over Glu-plasminogen. Similarly, Glu-plasminogen activation was augmented
by lysine or soluble fibrin. The ratio of the reaction rates of SC-UK and
TC-UK were comparable for Glu- and Lys-plasminogen. It is concluded that
there is a major difference in the catalytic activities of SC-UK and TC-UK
against plasminogen that is comparable to that against synthetic substrate.
Volume 69,
Issue 1,
pp. 22-26,
01/01/1987
Copyright © 1987 by The American Society of Hematology
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