Determination of dihedral Psi angles in large proteins by combining NH(N)/C(alpha)H(alpha) dipole/dipole cross-correlation and chemical shifts.
We propose a strategy based on the combination of experimental NH(N)/C(alpha)H(alpha) dipole/dipole cross-correlated relaxation rates and chemical shift analysis for the determination of Psi torsion angles in proteins. The method allows the determination of a dihedral angle that is not easily accessible by nuclear magnetic resonance (NMR). The measurement of dihedral angle restraints can be used for structure calculation, which is known to improve the quality of NMR structures. The method is of particular interest in the case of large proteins, for which spectral assignment of the nuclear Overhauser effect spectra, and therefore straightforward structural determination, is out of reach. One advantage of the method is that it is reasonably simple to implement, and could be used in association with other methods aiming at obtaining structural information on complex systems, such as residual dipolar coupling measurements. An illustrative example is analyzed in the case of the 30-kDa protein 6-phosphogluconolactonase.
Loth
Karine
K
Département de Chimie, Ecole Normale Supérieure, Paris, France.
Abergel
Daniel
D
Pelupessy
Philippe
P
Delarue
Marc
M
Lopes
Philippe
P
Ouazzani
Jamal
J
Duclert-Savatier
Nathalie
N
Nilges
Michael
M
Bodenhausen
Geoffrey
G
Stoven
Véronique
V
D000818:Animals; D002247:Carbon Isotopes; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry*; D018360:Crystallography, X-Ray; D006801:Humans; D008433:Mathematics; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D014346:Trypanosoma brucei brucei / Q000201:enzymology; D025801:Ubiquitin / Q000737:chemistry
D002247:Carbon Isotopes; D009587:Nitrogen Isotopes; D025801:Ubiquitin; D002265:Carboxylic Ester Hydrolases; C022363:6-phosphogluconolactonase