title: Expression in Escherichia coli, refolding, and purification of the recombinant mature form of human matrix metalloproteinase 7 (MMP-7).
abstract: In the latent pro-form of matrix metalloproteinase 7 (MMP-7), the cysteine residue in the pro-peptide binds the active-site zinc ion. Hence, recombinant active MMP-7 was prepared from pro-MMP-7 by modification of this cysteine residue with a mercuric reagent. In this study, mature MMP-7 was expressed in Escherichia coli as inclusion bodies, solubilized, and refolded with 1 M L-arginine. The purified product was indistinguishable from the one prepared from pro-MMP-7 as assessed by hydrolysis of (7-methoxycoumarin-4-yl)acetyl-L-Pro-L-Leu-Gly-L-Leu-[N(3)-(2,4-dinitrophenyl)-L-2,3-diaminopropionyl]-L-Ala-L-Arg-NH(2).
authors.affiliation: Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Kyoto, Japan.
authors.forename: Yuko
authors.initials: Y
authors.lastname: Muta
authors.forename: Natsuki
authors.initials: N
authors.lastname: Yasui
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Matsumiya
authors.forename: Motoki
authors.initials: M
authors.lastname: Kubo
authors.forename: Kuniyo
authors.initials: K
authors.lastname: Inouye
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000494:pharmacology; D001483:Base Sequence; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics*; D015870:Gene Expression; D006801:Humans; D006863:Hydrogen-Ion Concentration; D002479:Inclusion Bodies / Q000235:genetics; D020783:Matrix Metalloproteinase 7 / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D058849:Protein Refolding / Q000187:drug effects; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D012995:Solubility
chemical_list: D011994:Recombinant Proteins; D001120:Arginine; D020783:Matrix Metalloproteinase 7