Expression and purification of human matrilysin produced in baculovirus-infected insect cells.
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.
López de Turiso
J A
JA
Pharmacia Antibióticos Farma S.A., Research Department, Madrid, Spain.
Fernández
P
P
Barbacid
M M
MM
Mira
E
E
Quesada
A R
AR
Márquez
G
G
Aracil
M
M
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
D018076:DNA, Complementary; D011994:Recombinant Proteins; D008666:Metalloendopeptidases; D020783:Matrix Metalloproteinase 7