Extraction and partial characterization of a low molecular weight collagenase from normal rat kidney.
A matrix metalloproteinase with selective affinity for collagen was identified and partially purified from normal rat kidney. A two-step purification procedure consisting of gel filtration and affinity chromatography on Sepharose 4B-collagen resulted in an increase in specific activity of more than 3,000 times. The partially purified collagenase cleaved type I collagen and also showed low gelatinolytic activity. The enzyme required Ca2+ and revealed a neutral pH optimum; it was not inhibited by thiol or serineprotease inhibitors. Its activity was fully blocked using recombinant tissue inhibitor of metalloproteinases-1. Using SDS-PAGE and zymography, the estimated Mr of the collagenase was 16.5 x 10(3).
Vadillo-Ortega
F
F
Departamento de Bioquímica, Instituto Nacional de Perinatología, SS, México, D.F.
González-Avila
G
G
Montfort
I
I
Bermejo
L
L
Pérez-Tamayo
R
R
D000818:Animals; D002853:Chromatography, Liquid; D017364:Collagenases / Q000737:chemistry / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D051381:Rats; D017208:Rats, Wistar
D017364:Collagenases