Fast internal main-chain dynamics of human ubiquitin.
The fast internal dynamics of human ubiquitin have been studied by the analysis of 15N relaxation of backbone amide nitrogens. The amide 15N resonances have been assigned by use of heteronuclear multiple-quantum spectroscopy. Spin lattice relaxation times at 60.8 and 30.4 MHz and the steady-state nuclear Overhauser effect at 60.8 MHz have been determined for 67 amide 15N sites in the protein using two-dimensional spectroscopy. These data have been analyzed in terms of the model free treatment of Lipari and Szabo [Lipari, G., & Szabo, A. (1982) J. Am. Chem. Soc. 104, 4546-4559]. The global motion of the protein is shown to be isotropic and is characterized by a correlation time of 4.1 ns rad-1. The generalized order parameters (S2) of backbone amide N-H vectors in the globular region of the protein range from 0.5 to 0.95. No apparent correlation between secondary structure and generalized order parameters is observed. There is, however, a strong correlation between the magnitude of the generalized order parameters of a given N-H vector and the presence of hydrogen bonding of the amide hydrogen or its peptide bond associated carbonyl. Using a chemical shift tensor breadth of 160 ppm, the N-H vectors of peptide linkages participating in one or more hydrogen bonds to the main chain show an average generalized order parameter of 0.80 (SD 0.06), while those amide NH of peptide linkages free of hydrogen-bonding interactions with the main chain show an average order parameter of 0.69 (SD 0.06).(ABSTRACT TRUNCATED AT 250 WORDS)
Schneider
D M
DM
Department of Biochemistry, University of Illinois, Urbana, Illinois 61801.
Dellwo
M J
MJ
Wand
A J
AJ
D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D009587:Nitrogen Isotopes; D014452:Ubiquitins / Q000737:chemistry*
D009587:Nitrogen Isotopes; D014452:Ubiquitins