Identification of protein-protein interfaces by decreased amide proton solvent accessibility.

ABSTRACT: Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry was used to identify peptic fragments from protein complexes that retained deuterium under hydrogen exchange conditions due to decreased solvent accessibility at the interface of the complex. Short deuteration times allowed preferential labeling of rapidly exchanging surface amides so that primarily solvent accessibility changes and not conformational changes were detected. A single mass spectrum of the peptic digest mixture was analyzed to determine the deuterium content of all proteolytic fragments of the protein. The protein-protein interface was reliably indicated by those peptides that retained more deuterons in the complex compared with control experiments in which only one protein was present. The method wa

SUBMITTER: Mandell JG 

PROVIDER: S-EPMC24513 | biostudies-literature | 1998 Dec

REPOSITORIES: biostudies-literature