Elucidating in vivo structural dynamics in integral membrane protein by hydroxyl radical footprinting.

ABSTRACT: We describe here a novel footprinting technique to probe the in vivo structural dynamics of membrane protein. This method utilized in situ generation of hydroxyl radicals to oxidize and covalently modify biomolecules on living Escherichia coli cell surface. After enriching and purifying the membrane proteome, the modified amino acid residues of the protein were identified with tandem mass spectrometry to map the solvent-accessible surface of the protein that will form the footprint of in vivo structure of the protein. Of about 100 outer membrane proteins identified, we investigated the structure details of a typical beta-barrel structure, the porin OmpF. We found that six modified tryptic peptides of OmpF were reproducibly detected with 19 amino acids modified under the physiological condi

SUBMITTER: Zhu Y 

PROVIDER: S-EPMC2722762 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature