15N relaxation study of the cold shock protein CspB at various solvent viscosities.
For a detailed NMR study of the dynamics of the cold shock protein CspB from Bacillus subtilis, we determined (15)N transverse and longitudinal relaxation rates and heteronuclear nuclear Overhauser effects at different solvent viscosities. Up to a relative viscosity of 2, which is equivalent to 27% ethylene glycol (EG), the overall correlation time follows the linear Stokes-Einstein equation. At a relative viscosity of 6 (70% EG) the correlation time deviates from linearity by 30%, indicating that CspB tumbles at a higher rate as expected from the solvent viscosity probably due to a preferential binding of water molecules at the protein surface. The corresponding hydrodynamic radii, determined by NMR diffusion experiments, show no variation with viscosity. The amplitudes of intramolecular motions on a sub-nanosecond time scale revealed by an extended Lipari-Szabo analysis were mainly independent of the solvent viscosity. The lower limit of the NMR 'observation window' for the internal correlation time shifts above 0.5 ns at 70% EG, which is directly reflected in the experimentally derived internal correlation times. Chemical exchange contributions to the transverse relaxation rates derived from the Lipari-Szabo approach coincide with the experimentally determined values from the transverse (1)H-(15)N dipolar/(15)N chemical shift anisotropy relaxation interference. These contributions originate from fast protein folding reactions on a millisecond timescale, which get retarded at increased solvent viscosities.
Zeeb
Markus
M
Laboratorium für Biochemie, Universität Bayreuth, D-95440 Bayreuth, Germany.
Jacob
Maik H
MH
Schindler
Thomas
T
Balbach
Jochen
J
D001407:Bacillus / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry*; D041961:Deuterium Exchange Measurement; D008958:Models, Molecular; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D012997:Solvents / Q000737:chemistry; D013223:Statistics as Topic; D014783:Viscosity
D001426:Bacterial Proteins; D009587:Nitrogen Isotopes; D012997:Solvents; C077188:cold-shock protein CspB, Bacteria