A NMR experiment for simultaneous correlations of valine and leucine/isoleucine methyls with carbonyl chemical shifts in proteins

作者：Tugarinov Vitali^*; Venditti Vincenzo; Clore G Marius

来源：Journal of Biomolecular NMR, 2014, 58(1): 1-8.

DOI：10.1007/s10858-013-9803-1

• Chemical shift assignments
• Methyl labeling
• Enzyme I
• Methyl TROSY
• Isotope shifts

A methyl-detected 'out-and-back' NMR experiment for obtaining simultaneous correlations of methyl resonances of valine and isoleucine/leucine residues with backbone carbonyl chemical shifts, SIM-HMCM(CGCBCA)CO, is described. The developed pulse-scheme serves the purpose of convenience in recording a single data set for all Ile(delta 1), Leu(delta) and Val(gamma) (ILV) methyl positions instead of acquiring two separate spectra selective for valine or leucine/isoleucine residues. The SIM-HMCM(CGCBCA)CO experiment can be used for ILV methyl assignments in moderately sized protein systems (up to similar to 100 kDa) where the backbone chemical shifts of C-13(alpha), C-13(beta) and (CO)-C-13 are known from prior NMR studies and where some losses in sensitivity can be tolerated for the sake of an overall reduction in NMR acquisition time.

• 出版日期2014-1