Functional Production of Catalytic Domains of Human MMPs in Escherichia coli Periplasm

Methods Mol Biol. 2018:1731:65-72. doi: 10.1007/978-1-4939-7595-2_7.

Abstract

Due to their central roles in tumor growth and invasion, milligram-level amounts of active MMPs are frequently required for cancer research and development of chemical or biological MMP inhibitors. Here we describe methods for functional production of catalytic domains of MMPs (cdMMPs) in E. coli periplasm without refolding or activation process. We demonstrate applications of this straightforward approach for cdMMP-9, cdMMP-14, and cdMMP-14 mutants.

Keywords: Catalytic domain; MMP; Periplasmic expression.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Catalytic Domain / drug effects*
  • Drug Development / methods
  • Escherichia coli / cytology
  • Escherichia coli / metabolism*
  • Matrix Metalloproteinase 14 / genetics
  • Matrix Metalloproteinase 14 / isolation & purification*
  • Matrix Metalloproteinase 14 / metabolism
  • Matrix Metalloproteinase 9 / genetics
  • Matrix Metalloproteinase 9 / isolation & purification*
  • Matrix Metalloproteinase 9 / metabolism
  • Matrix Metalloproteinase Inhibitors / pharmacology
  • Mutation
  • Periplasm / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism

Substances

  • Matrix Metalloproteinase Inhibitors
  • Recombinant Proteins
  • MMP9 protein, human
  • Matrix Metalloproteinase 9
  • MMP14 protein, human
  • Matrix Metalloproteinase 14