¹³C-detected HN(CA)C and HMCMC experiments using a single methyl-reprotonated sample for unambiguous methyl resonance assignment

详细信息    查看全文

• 作者：Kaifeng Hu ; Beat V?geli and G. Marius Clore
• 关键词：13C ; detected NMR spectroscopy ; methyl resonance assignment ; IIBMannose
• 刊名：Journal of Biomolecular NMR
• 出版年：2006
• 出版时间：December, 2006
• 年：2006
• 卷：36
• 期：4
• 页码：259-266
• 全文大小：634 KB

文摘

Methyl groups provide an important source of structural and dynamic information in NMR studies of proteins and their complexes. For this purpose sequence-specific assignments of methyl 1H and 13C resonances are required. In this paper we propose the use of 13C-detected 3D HN(CA)C and HMCMC experiments for assignment of methyl 1H and 13C resonances using a single selectively methyl protonated, perdeuterated and 13C/15N-labeled sample. The high resolution afforded in the 13C directly-detected dimension allows one to rapidly and unambiguously establish correlations between backbone H_N strips from the 3D HN(CA)C spectrum and methyl group H_mC_m strips from the HMCMC spectrum by aligning all possible side-chain carbon chemical shifts and their multiplet splitting patterns. The applicability of these experiments for the assignment of methyl 1H and 13C resonances is demonstrated using the 18.6 kDa B domain of the Escherichia coli mannose transporter (IIBMannose).