Expression and characterization of human tissue inhibitor of metalloproteinases-1 in a baculovirus-insect cell system.
The baculovirus expression system was used to overexpress human tissue inhibitor of metalloproteinases-1 (TIMP-1). Approximately 5 mg of recombinant TIMP-1 was produced per 10(9) Sf9 insect cells infected with the recombinant baculovirus. The optimum time point for the production of biologically active rTIMP-1 was 20 hours postinfection. TIMP-1 activity was demonstrated by a soluble collagenase inhibition assay and reverse zymography. The baculovirus system has the advantage over previously described methods for generating human rTIMP-1 in that it generates large amounts of a fully glycosylated and active protein.
Gomez
D E
DE
Division of Cancer Etiology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
Lindsay
C K
CK
Cottam
D W
DW
Nason
A M
AM
Thorgeirsson
U P
UP
D000818:Animals; D016367:Baculoviridae; D015153:Blotting, Western; D002460:Cell Line; D003094:Collagen / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D005822:Genetic Vectors; D006023:Glycoproteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D006031:Glycosylation; D006801:Humans; D017403:In Situ Hybridization; D007700:Kinetics; D061965:Matrix Metalloproteinase Inhibitors; D008970:Molecular Weight; D009036:Moths; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D013997:Time Factors; D019714:Tissue Inhibitor of Metalloproteinases; D014162:Transfection
D006023:Glycoproteins; D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D003094:Collagen