Assignment of the aliphatic 1H and 13C resonances of the Bacillus subtilis glucose permease IIA domain using double- and triple-resonance heteronuclear three-dimensional NMR spectroscopy.
Assignment of the aliphatic 1H and 13C resonances of the Bacillus subtilis glucose permease IIA domain using double- and triple-resonance heteronuclear three-dimensional NMR spectroscopy.
复制标题
使用双共振和三共振异核三维 NMR 光谱分配枯草芽孢杆菌葡萄糖通透酶 IIA 结构域的脂肪族 1H 和 13C 共振。
DOI：
10.1021/bi00133a005
复制
发表时间：
1992
期刊：
影响因子：
2.9
通讯作者：
Wright,PE
中科院分区：
文献类型：
--
作者：
Fairbrother,WJ;Palmer3rd,AG;Rance,M;Reizer,J;SaierJr,MH;Wright,PE
斯克里普斯研究所分子生物学系，拉霍亚，加利福尼亚州92037，和加利福尼亚大学圣地亚哥分校生物学系，拉霍亚，加州92093-0116接收于1991年12月26日;修订版手册接收于1992年2月27日摘要：利用双共振和三共振三维（3D）NMR实验，对枯草芽孢杆菌IIAglc结构域的脂族和13 C共振进行了几乎完全的归属。使用恒定时间3D三重共振HCA（CO）N实验来获得13 C“共振的序列特异性分配，该实验将一个残基的”和13 C“化学位移与下一个残基的酰胺15 N化学位移相关联。先前已经主要使用3D-15 NOESY-HMQC和TOC-SY-HMQC实验顺序地指定了 *“和酰胺15 N化学位移[Fairbrother，W. J.，Cavanagh，J.，Dyson，H. J.，Palmer，A. G.，Ill，Sutrina，S. L.，Reizer，J.，赛尔，H、小的，& Wright，P. E.（1991）Biochemistry 30，6896-6907]。用3D HCCH-COSY和HCCH-TOCSY光谱鉴定了侧链自旋系统，并根据它们的π和13 C“化学位移对其进行了顺序归属。3D HCCH和HCA（CO）N实验依赖于大的杂原子单键J耦合的相干转移，因此提供了一个相当大的优势，比传统的'H-' H相关实验，依赖于-3 * J耦合，其中，对于蛋白质的大小IIA 8 ** 0（17.4 kDa），可能会显着小于线宽。本文报道的转让是必不可少的高分辨率的解决方案结构的IIAglc域的B的测定。使用3D和4D异源编辑的NOESY实验对枯草芽孢杆菌进行鉴定;这些鉴定已用于分析3D-15 NOESY-HMQC和H-13 C NOESY-HSQC光谱并计算低分辨率结构[Fairbrother，W. J.，Gippert，GP，Reizer，J.，赛尔，H、小的，& Wright，P. E. 04 The Dog of the Woman（1992）296，148-152]。
Department of Molecular Biology, The Scripps Research Institute, La Jolla, California 92037, and Department of Biology, University of California at San Diego, La Jolla, California 92093-0116 Received December 26, 1991; Revised Manuscript Received February 27, 1992 abstract: Nearly complete assignment of the aliphatic and 13C resonances of the IIAglc domain of Bacillus subtilis has been achievedusing a combination of double-and triple-resonance three-dimensional (3D) NMR experiments. A constant-time 3D triple-resonance HCA (CO) N experiment, which correlates the" and 13C “chemical shifts of one residue with the amide 15N chemical shift of the following residue, was used to obtain sequence-specific assignments of the 13C “resonances. The*“and amide 15N chemical shifts had been sequentially assigned previously using principally 3D-15 NOESY-HMQC and TOC-SY-HMQC experiments [Fairbrother, W. J., Cavanagh, J., Dyson, H. J., Palmer, A. G., Ill, Sutrina, S. L., Reizer, J., Saier,. H., Jr., & Wright, P. E.(1991) Biochemistry 30, 6896-6907]. The side-chain spin systems were identified using 3D HCCH-COSY and HCCH-TOCSY spectra and were assigned sequentially on the basis of their ß and 13C “chemical shifts. The 3D HCCH and HCA (CO) N experiments rely on large heteronuclear one-bond J couplings for coherence transfers and thereforeoffer a considerable advantage over conventional'H-'H correlation experiments that rely on-3**** J couplings, which, for proteins the size of IIA8** 0 (17.4 kDa), may be significantly smaller than the line widths. The assignments reported herein are essential for the determination of the high-resolution solution structure of the IIAglc domain of B. subtilis using 3D and 4D heteronuclear edited NOESY experiments; these assignments have been used to analyze 3D-15 NOESY-HMQC and'H-13C NOESY-HSQC spectra and calculate a low-resolution structure [Fairbrother, W. J., Gippert, GP, Reizer, J., Saier,. H., Jr., & Wright, P. E.(1992) FEBS Lett. 296, 148-152].