Secretion of single-chain urokinase-type plasminogen activator from insect cells.
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.
King
L A
LA
School of Biological and Molecular Sciences, Oxford Polytechnic, Headington, U.K.
Kaur
K
K
Mann
S G
SG
Lawrie
A M
AM
Steven
J
J
Ogden
J E
JE
D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D015153:Blotting, Western; D002478:Cells, Cultured; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D009036:Moths; D000077183:Occlusion Body Matrix Proteins; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics; D015678:Viral Structural Proteins
D000077183:Occlusion Body Matrix Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins; C009829:polyhedrin protein, Nucleopolyhedrovirus; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator