Production of a bioactive recombinant chicken matrix metalloproteinase-11 peptide in Escherichia coli.

Matrix metalloproteinase-11 (MMP-11) is known to be highly expressed in metastatic and most invasive forms of tumors. Being selectively expressed in tumor tissues MMP-11 is a promising target for immunotherapy against tumors. Here we report the production of a thioredoxin-tagged bioactive recombinant chicken MMP-11 peptide excluding the secretory signal and propeptide in Escherichia coli T7 Express lysY using pET32b(+) vector. High-level expression and purification of the bioactive peptide were achieved by induction with 1.0 mM IPTG for 4 H at 37°C followed by affinity chromatography under denaturing condition and slow dialysis. The recombinant peptide exhibited both caseinolytic and gelatinase activities without requiring activation by APMA. The antisera raised against the peptide in rabbit showed strong reaction with the whole recombinant peptide as well as the 37 kDa cMMP-11 mature peptide and cross-reactivity with a 43 kDa protein in murine breast tumor of 4T1 origin in Western blot analysis. The 43 kDa protein in the tumor homogenate showed immunoreactivity with a monoclonal antibody against human MMP-11 suggesting it to be murine MMP-11 having cross-reactivity with the antisera raised against chicken MMP-11 peptide. Altogether, the study characterized the production of a bioactive and immunogenic recombinant chicken MMP-11 peptide in E. coli This article is protected by copyright. All rights reserved