Cloning, expression, purification, and characterization of rat MMP-12

Citation
Jy. Fu et al., Cloning, expression, purification, and characterization of rat MMP-12, PROT EX PUR, 21(2), 2001, pp. 268-274
Citations number
16
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Biochemistry & Biophysics
Journal title
PROTEIN EXPRESSION AND PURIFICATION
ISSN journal
1046-5928 → ACNP
Volume
21
Issue
2
Year of publication
2001
Pages
268 - 274
Database
ISI
SICI code
1046-5928(200103)21:2<268:CEPACO>2.0.ZU;2-J
Abstract
Macrophage metalloelastase (MMP-12) is implicated in the pathology of many diseases such as emphysema, aortic lesions and cancer. Recently, MMP-12 was cloned and purified from mouse and human macrophages. We report here the e xpression of the full-length and catalytic domain of rat MMP-12 in Escheric hia coli and characterization of the purified enzyme. Inclusion bodies of e xpressed rat MMP-12 catalytic domain were denatured and refolded using a ne w method, and then affinity purified to near homogeneity with zinc chelatin g Sepharose, The purified rat MMP-18 catalytic domain was highly active in digesting substrates, having a K-m of 12 muM and optimal pH of 7.5-8.5. Dur ing investigation of natural substrate specificity, we found that rat MMP 1 2 catalytic domain was able to completely degrade collagen-V, partially deg rade collagen-I, but it was unable to digest collagen-IV. The enzyme could also degrade osteonectin, vitronectin, and fibronectin, but not laminin and albumin. The catalytic properties and natural substrate specificity of rat MMP-12 catalytic domain differed from those of human MMP-12 catalytic doma in, (C) 2001 Academic Press.