An optimized N^pro-based method for the expression and purification of intrinsically disordered proteins for an NMR study.

ABSTRACT: Intrinsically disordered proteins (IDPs) are an emerging concept. IDPs have high flexibility in their polypeptide chains, lacking a stable 3-dimensional structure. Because of the difficulty in performing X-ray crystallography for IDPs, nuclear magnetic resonance (NMR) spectroscopy is the first choice for atomic-level investigation of their nature. Given that isotopically labeled IDP samples are necessary for NMR study, a robust and cost-effective protocol for bacterial expression and purification of IDP is also needed. We employed the N^pro (EDDIE)-autoprotease fusion protein system. Although IDPs are believed to be readily degraded by endogenous proteases when expressed in Escherichia coli, N^pro-fused IDPs showed excellent resistance to degradation. Seven IDPs

SUBMITTER: Goda N 

PROVIDER: S-EPMC5314886 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature