Purification and properties of a small latent matrix metalloproteinase of the rat uterus.
A small metalloproteinase that digests Azocoll was found in the uterus of the rat. Its activity increased to high levels during the postpartum period in parallel with the breakdown of the extracellular matrix exclusive of collagen (Sellers, A., and Woessner, J.F., Jr. (1980) Biochem. J. 189, 521-531). This enzyme has now been purified almost 7,000-fold to homogeneity from 12 g of tissue using molecular sieve chromatography, blue sepharose chromatography, and zinc-chelate chromatography. Gel electrophoresis with sodium dodecyl sulfate and dithiothreitol gives Mr = 28,000 for the latent form of the enzyme and Mr = 19,000 for the active form that arises spontaneously or by treatment with aminophenylmercuric acetate. The enzyme digests components of the extracellular matrix including gelatins of types I, III, IV, and V, fibronectin, and proteoglycan. It digests the alpha 2(I) chain of gelatin in preference to the alpha 1(I) chain and cleaves dinitrophenyl-Pro-Leu-Gly-Ile-Ala-Gly-Pro-D-Arg. It cleaves the B chain of insulin at two points: Ala14-Leu15 and Tyr16-Leu17. It has no action on collagens of types I, III, IV, or V at 26 degrees C and no action on elastin or phenylazo-Pro-Leu-Gly-Pro-D-Arg. The pH optimum is at pH 7 and the pI at 5.9. The enzyme requires zinc and calcium ions for activity; cobalt and strontium can partially replace these metal ions. The enzyme is not inhibited by low levels of phosphoramidon or Zincov. Its properties clearly distinguish it from collagenase, gelatinase (matrix metalloproteinase 2), and stromelysin (matrix metalloproteinase 3); it therefore constitutes a further member of the family of extracellular matrix metalloendopeptidases. The name matrix metalloproteinase 7 is proposed.
Woessner
J F
JF
Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, Florida 33101.
Taplin
C J
CJ
D000818:Animals; D003094:Collagen / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000648:ultrastructure*; D005260:Female; D005780:Gelatin / Q000032:analysis; D007526:Isoelectric Point; D008666:Metalloendopeptidases / Q000302:isolation & purification*; D008970:Molecular Weight; D051381:Rats; D014599:Uterus / Q000201:enzymology*
D005780:Gelatin; D003094:Collagen; D008666:Metalloendopeptidases