The C-terminal domain (CTD) of MMP-2, which includes a hemopexin-like domain, has been increasingly studied as an alternative target in developing selective intervention strategies towards MMP-2. Moreover, The CTD itself has been implicated in a growing number of biological events, either MMP-dependent or –independent. The production of CTD, however, has been mostly based on the uncontrolled lysis of the latent ProMMP-2 or fusion protein expression that leaves a fusion tag. In this work we present a facile production of the untagged CTD in E. coli. The target protein was expressed as inclusion bodies, and we established an efficient wash and refolding strategy that allows us to obtain the target protein in extremely high purity. The yield was established at ~6 mg/L of the culture medium, which would greatly facilitate the production and hence the biological study of CTD. The method described herein might also prove useful for related (domain) proteins in MMP family and beyond.

越来越多地研究了MMP-2的C末端结构域（CTD），包括血红素样结构域，以此作为开发针对MMP-2的选择性干预策略的替代目标。此外，CTD本身与越来越多的MMP依赖性或非依赖性生物学事件有关。然而，CTD的产生主要基于潜伏的ProMMP-2的不受控制的裂解或留下融合标签的融合蛋白表达。在这项工作中，我们提出了在大肠杆菌中轻松生产未加标签的CTD的方法。目标蛋白表达为包涵体，我们建立了有效的洗涤和重折叠策略，使我们能够以极高的纯度获得目标蛋白。培养基的产量确定为〜6 mg / L，这将极大地促进CTD的生产和生物学研究。本文描述的方法也可能证明对MMP家族及以后的相关（域）蛋白有用。