Purification of aminophenyl mercuryacetate-activated human matrix metalloproteinase 1 and removal of the organomercurial in a single-step chromatography.
Matrix metalloproteinases are secreted from different cells as inactive zymogens. For their activation in vitro organomercurials may be used, the presence of which, however, can falsify activity assays and modulate the effects of the proteases in subsequent investigations. Here, we demonstrate the binding of human matrix metalloproteinase 1 to a thiophilic resin (mercaptoethylquinazolinedione derivatized agarose) and take advantage of this thiophilic interaction for the purification of organomercurial activated matrix metalloproteinase 1 from the supernatant of a thyroid carcinoma cell line in connection with the simultaneous removal of the activator.
Huse
K
K
Department of Internal Medicine III, University of Leipzig, Germany.
Wippich
P
P
Gutknecht
D
D
Aust
G
G
Scholz
G H
GH
D002846:Chromatography, Affinity / Q000379:methods; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D006801:Humans; D020781:Matrix Metalloproteinase 1 / Q000302:isolation & purification* / Q000378:metabolism*; D010662:Phenylmercuric Acetate / Q000031:analogs & derivatives* / Q000494:pharmacology; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology*; D013964:Thyroid Neoplasms / Q000201:enzymology; D014407:Tumor Cells, Cultured
D003470:Culture Media; D013439:Sulfhydryl Reagents; C010024:4-aminophenylmercuriacetate; D020781:Matrix Metalloproteinase 1; D010662:Phenylmercuric Acetate