Ikura M, Marion D, Kay L E, Shih H, Krinks M, Klee C B, Bax A
Laboratory of Chemical Physics, NIDDK, National Institute of Health, Bethesda, MD 20892.
Biochem Pharmacol. 1990 Jul 1;40(1):153-60. doi: 10.1016/0006-2952(90)90190-v.
New methods are described that permit detailed analysis of the NMR spectra of calmodulin, an alpha-helical protein with a molecular weight of 16.7 kD. Two complementary approaches have been used: uniform labeling with 15N and labeling of specific amino acids with either 15N or 13C. It is demonstrated that uniform 15N labeling permits the recording of sensitive three-dimensional (3D) NMR spectra that show far better resolution than their conventional two-dimensional analogs. Selective 15N labeling of amino acids can be used for identifying the type of amino acid, providing information that is essential for the analysis of the 3D spectra. Simultaneous selective labeling with both 15N and 13C can provide a number of unique backbone assignments from which sequential assignment can be continued.
本文描述了一些新方法，这些方法可用于对钙调蛋白的核磁共振谱进行详细分析，钙调蛋白是一种分子量为16.7 kD的α螺旋蛋白。采用了两种互补的方法：用15N进行均匀标记以及用15N或13C对特定氨基酸进行标记。结果表明，均匀的15N标记能够记录灵敏的三维（3D）核磁共振谱，其分辨率远高于传统的二维类似谱。对氨基酸进行选择性15N标记可用于识别氨基酸类型，为3D谱分析提供至关重要的信息。同时用15N和13C进行选择性标记可提供许多独特的主链归属，据此可继续进行序列归属。
