Relaxation-optimized NMR spectroscopy of methylene groups in proteins and nucleic acids

被引:69
|
作者
Miclet, E [1 ]
Williams, DC [1 ]
Clore, GM [1 ]
Bryce, DL [1 ]
Boisbouvier, J [1 ]
Bax, A [1 ]
机构
[1] NIDDKD, Phys Chem Lab, Natl Inst Hlth, Bethesda, MD 20892 USA
关键词
D O I
10.1021/ja047904v
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
A large fraction of hydrogens in proteins and nucleic acids is of the methylene type. Their detailed study, however, in terms of structure and dynamics by NMR spectroscopy is hampered by their fast relaxation properties, which give rise to low sensitivity and resolution. It is demonstrated that six different relaxation interference processes, involving H-1-C-13 and H-1-H-1 dipolar interactions and H-1 and C-13 chemical shift anisotropy, can be used simultaneously to mitigate these problems effectively. The approach is applicable to the majority of NMR experiments commonly used to study side chain and backbone conformation. For proteins, its efficiency is evaluated quantitatively for two samples: the third IgG-bincling domain from Streptococcal Protein G and the protein calmodulin complexed with a 26-residue target peptide. Gains in both resolution and sensitivity by up to factors of 3.2 and 2.0, respectively, are observed for Gly residues at high magnetic field strengths, but even at much lower fields gains remain substantial. The resolution enhancement obtained for methylene groups makes possible a detailed analysis of spectral regions commonly considered inaccessible due to spectral crowding. For DNA, the high resolution now obtainable for C-5' sites permits an H-5'/H-5"-based sequential NOE assignment procedure, complementary to the conventional base-H-1'/H-2'/H-2" pathway.
引用
收藏
页码:10560 / 10570
页数:11
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