title: Expression and characterization of human tissue inhibitor of metalloproteinases-1 in a baculovirus-insect cell system.
abstract: 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.
authors.affiliation: Division of Cancer Etiology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: D E
authors.initials: DE
authors.lastname: Gomez
authors.forename: C K
authors.initials: CK
authors.lastname: Lindsay
authors.forename: D W
authors.initials: DW
authors.lastname: Cottam
authors.forename: A M
authors.initials: AM
authors.lastname: Nason
authors.forename: U P
authors.initials: UP
authors.lastname: Thorgeirsson
mesh_terms: 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
chemical_list: D006023:Glycoproteins; D061965:Matrix Metalloproteinase Inhibitors; D011994:Recombinant Proteins; D019714:Tissue Inhibitor of Metalloproteinases; D003094:Collagen