A study of protein side-chain dynamics from new 2H auto-correlation and 13C cross-correlation NMR experiments: application to the N-terminal SH3 domain from drk.
Two new NMR experiments are presented for measuring side-chain dynamics in proteins. The first method, requiring 15N, 13C, approximately 50% 2H-labeled protein, measures 2H T1 and T1p spin relaxation times at side-chain positions. A second experiment permits the straightforward measurement of 13C-1H dipole-dipole cross-correlation relaxation rates at 13C beta positions in 15N, 13C-labeled molecules. An excellent correlation is observed between order parameters, describing the amplitude of motion at these sites, obtained on the basis of 2H relaxation and dipole-dipole cross-correlation relaxation rates. Together these experiments provide a powerful approach for selecting appropriate motional models. The methods are applied to study the side-chain motional properties of the N-terminal SH3 domain from the signaling protein drk.
Yang
D
D
Department of Medical Genetics, University of Toronto, Ontario, Canada.
Mittermaier
A
A
Mok
Y K
YK
Kay
L E
LE
D000818:Animals; D002247:Carbon Isotopes; D003903:Deuterium; D029721:Drosophila Proteins; D019476:Insect Proteins / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D013816:Thermodynamics; D018909:src Homology Domains
D002247:Carbon Isotopes; D029721:Drosophila Proteins; D019476:Insect Proteins; C080530:drk protein, Drosophila; D003903:Deuterium