Anisotropic molecular rotational diffusion in 15N spin relaxation studies of protein mobility.
The backbone dynamics of the uniformly 15N-labeled N-terminal 63-residue DNA-binding domain of the 434 repressor has been characterized by measurements of the individual 15N longitudinal relaxation times, T1, transverse relaxation times, T2, and heteronuclear 15N[1H]-NOEs at 1H resonance frequencies of 400 and 750 MHz. The dependence of an apparent spherical top correlation time, tauR, on the orientation of the N-H bond vector with respect to the principal axes of the global diffusion tensor of the protein was used to establish the fact that the degree of anisotropy of the global molecular tumbling amounts to 1.2, which is in good agreement with the values obtained from model calculations of the hydrodynamic properties. A model-free analysis showed that even this small anisotropy leads to the implication of artifactual slow internal motions for at least two residues when the assumption of isotropic global motion is used. Additional residues may actually undergo internal motions on the same time scale as the global rotational diffusion, in which case the model-free approach would, however, be inappropriate for quantifying the correlation times and order parameters. Overall, the experiments with 434(1-63) demonstrate that the assumption of isotropic rotational reorientation may result in artifacts of model-free interpretations of spin relaxation data even for proteins with small deviations from spherical shape.
Luginbühl
P
P
Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule-Hönggerberg, Zürich, Switzerland.
Pervushin
K V
KV
Iwai
H
H
Wüthrich
K
K
D001665:Binding Sites; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D015394:Molecular Structure; D011487:Protein Conformation; D011994:Recombinant Proteins; D012097:Repressor Proteins / Q000737:chemistry* / Q000378:metabolism; D012996:Solutions; D014764:Viral Proteins
C063367:434-repressor protein, Bacteriophage 434; D011994:Recombinant Proteins; D012097:Repressor Proteins; D012996:Solutions; D014764:Viral Proteins; D004247:DNA