Biomaterial Database

Resolution of TIMP-free and TIMP-complexed 70kDa progelatinase from culture medium of Rous sarcoma virus-transformed chicken embryo fibroblasts. 1993

R T Aimes, and S M Nielsen-Preiss, and J P Quigley

Department of Biochemistry, State University of New York, Stony Brook 11794.

Chicken embryo fibroblasts (CEF) produce a 70kDa progelatinase, a member of the matrix metalloproteinase family, and secrete elevated levels of the enzyme upon transformation by Rous sarcoma virus (RSV). This enzyme can be purified by affinity chromatography complexed with a 21kDa tissue inhibitor of metalloproteinases (TIMP)-like molecule. Gel-filtration of the purified progelatinase suggests the presence of a mixed population of enzyme: a TIMP-complexed and a TIMP-free progelatinase. These two species were separated by Mono Q FPLC in the absence of denaturants. Quantitation of the purified progelatinase reveals that the transformed RSVCEF produce more TIMP-free enzyme than the normal CEF. Native PAGE analysis indicates that purified TIMP-free progelatinase is capable of binding to TIMP and generating a TIMP-complexed progelatinase. Treatment of the TIMP-free gelatinase with organomercurials results in a rapid conversion to the active 62kDa species and indicates that the TIMP-free progelatinase is more susceptible to activation than the TIMP-complexed progelatinase.

UI	MeSH Term	Description	Entries
D008666	Metalloendopeptidases	ENDOPEPTIDASES which use a metal such as ZINC in the catalytic mechanism.	Metallo-Endoproteinases,Metalloendopeptidase
D002461	Cell Line, Transformed	Eukaryotic cell line obtained in a quiescent or stationary phase which undergoes conversion to a state of unregulated growth in culture, resembling an in vitro tumor. It occurs spontaneously or through interaction with viruses, oncogenes, radiation, or drugs/chemicals.	Transformed Cell Line,Cell Lines, Transformed,Transformed Cell Lines
D002642	Chick Embryo	The developmental entity of a fertilized chicken egg (ZYGOTE). The developmental process begins about 24 h before the egg is laid at the BLASTODISC, a small whitish spot on the surface of the EGG YOLK. After 21 days of incubation, the embryo is fully developed before hatching.	Embryo, Chick,Chick Embryos,Embryos, Chick
D002850	Chromatography, Gel	Chromatography on non-ionic gels without regard to the mechanism of solute discrimination.	Chromatography, Exclusion,Chromatography, Gel Permeation,Chromatography, Molecular Sieve,Gel Filtration,Gel Filtration Chromatography,Chromatography, Size Exclusion,Exclusion Chromatography,Gel Chromatography,Gel Permeation Chromatography,Molecular Sieve Chromatography,Chromatography, Gel Filtration,Exclusion Chromatography, Size,Filtration Chromatography, Gel,Filtration, Gel,Sieve Chromatography, Molecular,Size Exclusion Chromatography
D002853	Chromatography, Liquid	Chromatographic techniques in which the mobile phase is a liquid.	Liquid Chromatography
D003470	Culture Media	Any liquid or solid preparation made specifically for the growth, storage, or transport of microorganisms or other types of cells. The variety of media that exist allow for the culturing of specific microorganisms and cell types, such as differential media, selective media, test media, and defined media. Solid media consist of liquid media that have been solidified with an agent such as AGAR or GELATIN.	Media, Culture
D004591	Electrophoresis, Polyacrylamide Gel	Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.	Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D004792	Enzyme Precursors	Physiologically inactive substances that can be converted to active enzymes.	Enzyme Precursor,Proenzyme,Proenzymes,Zymogen,Zymogens,Precursor, Enzyme,Precursors, Enzyme
D005109	Extracellular Matrix	A meshwork-like substance found within the extracellular space and in association with the basement membrane of the cell surface. It promotes cellular proliferation and provides a supporting structure to which cells or cell lysates in culture dishes adhere.	Matrix, Extracellular,Extracellular Matrices,Matrices, Extracellular
D005347	Fibroblasts	Connective tissue cells which secrete an extracellular matrix rich in collagen and other macromolecules.	Fibroblast