title: Observation of α-Helical Hydrogen-Bond Cooperativity in an Intact Protein.
abstract: The presence and extent of hydrogen-bonding (H-bonding) cooperativity in proteins remains a fundamental question, which in the past has been studied extensively, mostly by infrared and fluorescence measurements on model peptides. We demonstrate that such cooperativity can be studied in an intact protein by hydrogen/deuterium exchange NMR spectroscopy. The method is based on the fact that substitution of NH by ND in a backbone amide group slightly weakens the N-H···O═C hydrogen bond. Our results show that such substitution at position i in an α-helix impacts the (1)H and (15)N chemical shifts of the amide sites of residues i - 3 to i + 3. Quantum mechanical calculations indicate that the upfield shifts of (1)H and (15)N resonances at site i, observed upon H/D exchanges at sites i - 3, i + 1, i + 2, and i + 3, correspond to a decrease of the ith backbone amide electric dipole moment, which weakens its H-bonding and long-range electrostatic interactions with other backbone amides in the α-helix. These results provide new quantitative insights into the cooperativity of H-bonding in protein α-helices.
authors: Jingwen
authors: J
authors: Li
authors: Yefei
authors: Y
authors: Wang
authors: Jingfei
authors: J
authors: Chen
authors: National Center for Protein Science Shanghai, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences , Shanghai 201210, China.
authors: Zhijun
authors: Z
authors: Liu
authors: Laboratory of Chemical Physics, NIDDK, National Institutes of Health , Bethesda, Maryland 20892-0520, United States.
authors: Ad
authors: A
authors: Bax
authors: Lishan
authors: L
authors: Yao
mesh_terms: D006860:Hydrogen Bonding; D019906:Nuclear Magnetic Resonance, Biomolecular; D011506:Proteins / Q000737:chemistry*
chemical_list: D011506:Proteins