Probing the Impact of Molecular Chaperones on the Refoldability of the E. coli Proteome

Abstract

We have demonstrated that roughly half of the E. coli proteome is unable to refold to their native structures following denaturation with chemical denaturants. To accomplish these studies, we devised a limited proteolysis mass-spectrometry (LiP-MS) approach, in which a permissive protease that cleaves at flexible regions is used to probe the structural differences between the native and refolded forms of proteins in whole lysates. Here, we interrogate the ability of several molecular chaperone systems (trigger factor, HSP70, and the chaperonin GroEL/GroES) to rescue the refoldability of proteins that cannot refold on their own.