title: Expression and purification of human matrilysin produced in baculovirus-infected insect cells.
abstract: The baculovirus expression system was used to produce recombinant human matrilysin. Expression of promatrilysin reached a peak at 72 h post-infection. Most of the recombinant protein remained in the intracellular fraction in an insoluble form, which after renaturation was purified by S-Sepharose and Green A Dyematrex chromatography in order to remove host proteases. Active recombinant matrilysin degraded casein, type I and type IV collagens and fibronectin. Expression of recombinant human matrilysin using the baculovirus system represents a useful tool for obtaining large amounts of this metalloproteinase in order to carry out further biochemical studies and to screen for inhibitors.
authors.affiliation: Pharmacia Antibióticos Farma S.A., Research Department, Madrid, Spain.
authors.forename: J A
authors.initials: JA
authors.lastname: López de Turiso
authors.forename: P
authors.initials: P
authors.lastname: Fernández
authors.forename: M M
authors.initials: MM
authors.lastname: Barbacid
authors.forename: E
authors.initials: E
authors.lastname: Mira
authors.forename: A R
authors.initials: AR
authors.lastname: Quesada
authors.forename: G
authors.initials: G
authors.lastname: Márquez
authors.forename: M
authors.initials: M
authors.lastname: Aracil
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D001483:Base Sequence; D002460:Cell Line; D003110:Colonic Neoplasms; D018076:DNA, Complementary / Q000737:chemistry; D015870:Gene Expression; D006801:Humans; D020783:Matrix Metalloproteinase 7; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D018411:Spodoptera; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D008666:Metalloendopeptidases; D020783:Matrix Metalloproteinase 7