Protein dynamics measurements by TROSY-based NMR experiments.
The described TROSY-based experiments for investigating backbone dynamics of proteins make it possible to elucidate internal motions in large proteins via measurements of T(1), T(2), and NOE of backbone (15)N nuclei. In our proposed sequences, the INEPT sequence is eliminated and the PEP sequence is replaced by the ST2-PT sequence from the HSQC-based experiments. This has the benefit of shortening the pulse sequences by 5.4 ms (=1/2J) and results in an increase in the intrinsic sensitivity of the proposed TROSY-based experiments. The TROSY-based experiments are on average of 13% more sensitive than the corresponding HSQC-based experiments on a uniformly (15)N-labeled Xenopus laevis calcium-bound calmodulin sample on a 750-MHz spectrometer at 5 degrees C. The amide proton linewidths of the TROSY-based experiments are 2-13 Hz narrower than those of the HSQC experiments. More sensitivity gain and higher resolution are expected if the protein sample is deuterated.
Zhu
G
G
Department of Biochemistry, The Hong Kong University of Science and Technology, Kowloon, Hong Kong. gzhu@ust.hk
Xia
Y
Y
Nicholson
L K
LK
Sze
K H
KH
D000577:Amides; D000818:Animals; D002147:Calmodulin / Q000737:chemistry*; D003080:Cold Temperature; D003903:Deuterium; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D009587:Nitrogen Isotopes; D011487:Protein Conformation; D012680:Sensitivity and Specificity; D013997:Time Factors; D014982:Xenopus laevis
D000577:Amides; D002147:Calmodulin; D009587:Nitrogen Isotopes; D003903:Deuterium