Abstract
A two-dimensional TROSY-based SIM-13Cm–1Hm/1H–15N NMR experiment for simultaneous measurements of methyl 1 D CH and backbone amide 1 D NH residual dipolar couplings (RDC) in {U-[15N,2H]; Ileδ1-[13CH3]; Leu,Val-[13CH3/12CD3]}-labeled samples of large proteins is described. Significant variation in the alignment tensor of the 82-kDa enzyme Malate synthase G is observed as a function of only slight changes in experimental conditions. The SIM-13Cm–1Hm/1H–15N data sets provide convenient means of establishing the alignment tensor characteristics via the measurement of 1 D NH RDCs in the same protein sample.
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Acknowledgments
The authors thank Prof. W.-Y. Choy (University of Western Ontario, Canada) for the program RDCA2.0 used to fit and visualize alignment tensors derived from 1 D NH couplings, and Prof. Joel Tolman (Johns Hopkins University, Maryland) for useful discussions.
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Xinli Liao is on leave from the Department of Chemistry, Xiamen University, China.
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Liao, X., Godoy-Ruiz, R., Guo, C. et al. Simultaneous measurement of 1H–15N and Methyl 1Hm–13Cm residual dipolar couplings in large proteins. J Biomol NMR 51, 191 (2011). https://doi.org/10.1007/s10858-011-9553-x
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DOI: https://doi.org/10.1007/s10858-011-9553-x