title: Method for isolation of MMP-7-ase isoenzymes from rat muscle tissue extract.
abstract: Two MMP-7-ase isoenzymes were purified 100-fold from rat muscle extract to apparent homogeneity, with an overall yield of 10%, using homogenization, ultracentrifugation, high-performance aqueous size-exclusion and high-performance anion exchange chromatography methods. When using a TSK G-2000SW column, the separation resulted in a 6-fold purification and 30% recovery of isoenzymes B and C. This concentrated enzyme extract was then passed through a TSK-DEAE-2SW column, using salt gradient at pH 7.5, with an additional 25-fold purification and 90% recovery of the isoenzymes. Two symmetrical enzyme peaks, representing isoenzymes B and C, were detected when performing purity tests of the active enzymes on the anion exchanger and reversed-phase HPLC columns. The procedures involved are extraction, ultracentrifugation, chromatographies and enzyme assays and require less than five hours.
authors.affiliation: Endocrine, Polypeptide and Cancer Institute, Veterans Administration Medical Center, New Orleans, Louisiana 70146.
authors.forename: J G
authors.initials: JG
authors.lastname: Guoth
authors.forename: I
authors.initials: I
authors.lastname: Sohar
authors.forename: M
authors.initials: M
authors.lastname: Zarandi
authors.forename: T
authors.initials: T
authors.lastname: Janaky
authors.forename: A V
authors.initials: AV
authors.lastname: Schally
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007527:Isoenzymes / Q000302:isolation & purification*; D008666:Metalloendopeptidases / Q000302:isolation & purification*; D009132:Muscles / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D007527:Isoenzymes; D008666:Metalloendopeptidases