Purification of a procollagenase-activator present in medium of cultured guinea pig carrageenin granuloma.
Activation of procollagenase constitutes a crucial event in collagenolytic activity regulation. In this study we have purified by DEAE-cellulose, Ultrogel AcA-44, and zinc chelate sepharose chromatographies, a procollagenase-activator from the culture medium of the guinea pig carrageenin granuloma model. On SDS-PAGE, the activator migrates as a principal band of Mr approximately 44,000. The molecule activates procollagenase from human lung fibroblasts in a concentration dependent manner and an enhancement of collagenase activity of trypsin-treated crude culture medium was observed. A loss of about 50% of its activity occurs after heating. In addition, this activator degrades gelatin and casein. All these data suggest that this procollagenase-activator might be stromelysin. The activator was found in both phases of the granuloma, at 7 days when collagen is actively deposited and an important proportion of the collagenolytic activity remains in latent form; and at 14 days, when this enzymatic activity is fully expressed.
Pardo
A
A
Facultad de Ciencias, Universidad Nacional Autonoma de Mexico, D.F.
Ramirez
R
R
Gutierrez-Kobeh
L
L
Mendoza
F
F
Bauer
E
E
Selman
M
M
D000818:Animals; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003094:Collagen / Q000378:metabolism*; D017364:Collagenases; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D006099:Granuloma / Q000139:chemically induced / Q000201:enzymology; D006168:Guinea Pigs; D019278:Matrix Metalloproteinase 3; D008666:Metalloendopeptidases / Q000302:isolation & purification*; D003012:Microbial Collagenase / Q000378:metabolism*; D012867:Skin / Q000201:enzymology; D015032:Zinc
D004792:Enzyme Precursors; D003094:Collagen; D017364:Collagenases; D008666:Metalloendopeptidases; C022440:procollagenase; D019278:Matrix Metalloproteinase 3; D003012:Microbial Collagenase; D015032:Zinc