Direct production of functional matrix metalloproteinase--14 without refolding or activation and its application for in vitro inhibition assays.
Biotechnol Bioeng
; 113(4): 717-23, 2016 Apr.
Article
en En
| MEDLINE
| ID: mdl-26416249
ABSTRACT
Human matrix metalloproteinase (MMP)-14, a membrane-bound zinc endopeptidase, is one of the most important cancer targets because it plays central roles in tumor growth and invasion. Large amounts of active MMP-14 are required for cancer research and the development of chemical or biological MMP-14 inhibitors. Current methods of MMP-14 production through refolding and activation are labor-intensive, time-consuming, and often associated with low recovery rates, lot-to-lot variation and heterogeneous products. Here, we report direct production of the catalytic domain of MMP-14 in the periplasmic space of Escherichia coli. 0.5 mg/L of functional MMP-14 was produced without tedious refolding or problematic activation process. MMP-14 prepared by simple periplasmic treatment can be readily utilized to evaluate the potencies of chemical and antibody-based inhibitors. Furthermore, co-expression of both MMP-14 and antibody Fab fragments in the periplasm facilitated inhibitory antibody screening by avoiding purification of MMP-14 or Fabs. We expect this MMP-14 expression strategy can expedite the development of therapeutic drugs targeting MMPs with biological significance.
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Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Proteínas Recombinantes
/
Escherichia coli
/
Metaloproteinasa 14 de la Matriz
Límite:
Humans
Idioma:
En
Revista:
Biotechnol Bioeng
Año:
2016
Tipo del documento:
Article