TROSY-type triple-resonance experiments for sequential NMR assignments of large proteins
TROSY-type triple-resonance experiments for sequential NMR assignments of large proteins
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DOI：
10.1021/ja9834226
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发表时间：
1999-02-03
影响因子：
15
通讯作者：
Wüthrich, K
中科院分区：
文献类型：
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作者：
Salzmann, M;Wider, G;Wüthrich, K
对四个三重共振实验[N-15，H-1]-TROSY-HN(CO)CA、[N-15，H-1]-TROSY-HN(CA)CO、[N-15，H-1]-TROSY-HNCACB和[N-15，H-1]-TROSY-HN(CO)CACB进行了横向弛豫优化光谱(TROSY)研究。与[N-15，H-1]-TROSY-HNCA和[N-15，H-1]-TROSY-HNCO(Salzmann，M.；Pervushin，K.；Wide，G.；Senn，H.Wothrich，K.proc.娜塔莉。阿卡德。SCI。美国1998年，13585-13590)这些实验代表了一套TROSY型三重共振实验，它使得蛋白质的顺序骨架指定成为可能。当与23 kDa H-2/C-13/N-15标记的蛋白质旋转酶23B一起使用时，与相应的传统核磁共振实验相比，平均而言，在整个氨基酸序列上，四个实验中的每一个的灵敏度都提高了3倍。因此，在三重共振实验中使用TROSY原理有望实现对比目前相应的传统核磁共振实验所能实现的更大蛋白质的共振指定。
Transverse relaxation-optimized spectroscopy (TROSY) was implemented in the four triple resonance experiments [N-15,H-1]-TROSY-HN(CO)CA, [N-15,H-1]-TROSY-HN(CA)CO, [N-15,H-1]-TROSY-HNCACB, and [N-15, H-1]-TROSY-HN(CO)CACB. Combined with [N-15,H-1]-TROSY-HNCA and [N-15,H-1]-TROSY-HNCO (Salzmann, M.; Pervushin, K.; Wider, G.; Senn, H. Wuthrich, K. Proc. Natl. Acad. Sci. U.S.A. 1998, 13585-13590) these experiments represent a suite of TROSY-type triple resonance experiments that enables sequential backbone assignment of proteins. When used with the 23 kDa H-2/C-13/N-15-labeled protein gyrase 23B, a comparison with the corresponding conventional NMR experiments showed, on average over the entire amino acid sequence, a 3-fold sensitivity gain for each of the four experiments. The use of the TROSY principle in triple resonance experiments thus promises to enable resonance assignments for significantly larger proteins than what is achievable today with the corresponding conventional NMR experiments.