Secretion of single-chain urokinase-type plasminogen activator from insect cells. |
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Authors: | L A King K Kaur S G Mann A M Lawrie J Steven J E Ogden |
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Institution: | School of Biological and Molecular Sciences, Oxford Polytechnic, Headington, U.K. |
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Abstract: | A cDNA encoding human urokinase-type plasminogen activator was inserted downstream from the polyhedrin promoter of the baculovirus Autographa californica nuclear polyhedrosis virus. A protein of similar Mr to urokinase (UK) was synthesized and approx. 90% was secreted from recombinant virus-infected Spodoptera frugiperda cells. Zymography and Western blotting analysis of the insect-derived protein demonstrated that it was comprised solely of the high-Mr form of UK. No low-Mr UK was detected. Amidolytic activity assays showed that 96% of the insect cell-derived UK was in the single-chain proenzyme form. The yield of UK from insect cells was 1986 international units/ml/10(6) infected cells. |
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