HRMAS: 1H-15N TROSY of fractionally deuterated (mostly at Calpha) KcsA

Citation

M. Czisch & R. Boelens, J. Magn. Reson. 134, 158-160 (1998) https://www.sciencedirect.com/science/article/abs/pii/S1090780798914835 K. Pervushin, G. Wider & K. Wuethrich, J. Biomol. NMR 12, 345-348 (1998) https://www.ncbi.nlm.nih.gov/pubmed/21136330 J. Weigelt, J. Am. Chem. Soc. 120, 10778-10779 (1998) https://pubs.acs.org/doi/abs/10.1021/ja982649y M. Rance, J.P. Loria & A.G. Palmer III, J. Magn. Reson. 136, 91-101 (1999) https://www.ncbi.nlm.nih.gov/pubmed/9887294 G. Zhu, X.M. Kong & K.H. Sze, J. Biomol. NMR 13, 77-81 (1999) https://www.ncbi.nlm.nih.gov/pubmed/21080266

Protocol

a. Increasing spinning frequency above 8kHz doesn't bring any improvements in most cases. b. SMSQ10.100 and SMSQ10.50 have better performance than SINE.100 and SINE.50 c. use -DLABEL_CN flag on with Crp80,0.5,20.1 pulse for 13C-15N labeled sample. The pulse refocuses 13C(O) and sharpens up 15N lines substantially

Samples

The experiments were performed with a precipitated 2H (Capha), U-13C/15N KcsA, LPR =1, 9:1 DOPE-DOPS 20% D2O citrate buffer pH 3.

Processing and analysis

Pulse Sequence and Parameter Set