Expression of human membrane type 1 matrix metalloproteinase in Pichia pastoris

Citation
M. Roderfeld et al., Expression of human membrane type 1 matrix metalloproteinase in Pichia pastoris, PROT EX PUR, 19(3), 2000, pp. 369-374
Citations number
40
Categorie Soggetti
Biochemistry & Biophysics
Journal title
PROTEIN EXPRESSION AND PURIFICATION
ISSN journal
10465928 → ACNP
Volume
19
Issue
3
Year of publication
2000
Pages
369 - 374
Database
ISI
SICI code
1046-5928(200008)19:3<369:EOHMT1>2.0.ZU;2-S
Abstract
A soluble, C-terminal truncated form of human membrane type 1 matrix metall oproteinase (MT1-MMP) containing the hemopexin-like domain was expressed in Pichia pastoris strain KM71. High levels of secreted protein were detected . Although the c-DNA for the proenzyme (Ala(21)-Glu(523) called Delta TM-MT 1-MMP) was cloned, almost only active MT1-MMP (Tyr(112)-GlU(523)) with iden tical N-terminus as described for the wildtype enzyme was isolated. This ac tive enzyme was highly purified and characterized with respect to its bioch emical properties, The recombinant protein showed high stability against au tolysis and proteolysis by yeast proteases, although the calculated in vivo half-life is rather low. The biochemical properties of this new MT1-MMP sp ecies were compared with the well-characterized catalytic domain (Ile(114)- Ile(318)) of MT1-MMP, The novel form of MT1-MMP exhibited a higher stabilit y against autolysis than the isolated catalytic domain (Ile(114)-Ile(318)). (C) 2000AcademicPress.