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Hydrogen-deuterium exchange strategy for delineation of contact sites in protein complexes.

ABSTRACT: We use NMR spectra to determine protein-protein contact sites by observing differences in amide proton hydrogen-deuterium exchange in the complex compared to the free protein in solution. Aprotic organic solvents are used to preserve H/D labeling patterns that would be scrambled in water solutions. The binding site between the mammalian co-chaperone Aha1 with the middle domain of the chaperone Hsp90 obtained by our H/D exchange method corresponds well with that in the X-ray crystal structure of the homologous complex from yeast, even to the observation of a secondary binding site. This method can potentially provide data for complexes with unknown structure and for large or dynamic complexes inaccessible via NMR and X-ray methods.

SUBMITTER: Dyson HJ

PROVIDER: S-EPMC2440508 | biostudies-literature | 2008 Apr

REPOSITORIES: biostudies-literature

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Hydrogen-deuterium exchange strategy for delineation of contact sites in protein complexes.

Dyson H Jane HJ Kostic Milka M Liu Jeff J Martinez-Yamout Maria A MA

FEBS letters 20080407 10

We use NMR spectra to determine protein-protein contact sites by observing differences in amide proton hydrogen-deuterium exchange in the complex compared to the free protein in solution. Aprotic organic solvents are used to preserve H/D labeling patterns that would be scrambled in water solutions. The binding site between the mammalian co-chaperone Aha1 with the middle domain of the chaperone Hsp90 obtained by our H/D exchange method corresponds well with that in the X-ray crystal structure of ...[more]

PMID: 18396165

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