Steric trapping reveals a cooperativity network in the intramembrane protease GlpG.

ABSTRACT: Membrane proteins are assembled through balanced interactions among proteins, lipids and water. Studying their folding while maintaining the native lipid environment is necessary but challenging. Here we present methods for analyzing key elements of membrane protein folding including thermodynamic stability, compactness of the unfolded state and folding cooperativity under native conditions. The methods are based on steric trapping, which couples the unfolding of a doubly biotinylated protein to the binding of monovalent streptavidin (mSA). We further advanced this technology for general application by developing versatile biotin probes possessing spectroscopic reporters that are sensitized by mSA binding or protein unfolding. By applying these methods to the Escherichia coli intramembrane

SUBMITTER: Guo R 

PROVIDER: S-EPMC4837050 | biostudies-literature | 2016 May

REPOSITORIES: biostudies-literature