Sequence-specific 1H NMR resonance assignments of Bacillus subtilis HPr: use of spectra obtained from mutants to resolve spectral overlap.
Sequence-specific 1H NMR resonance assignments of Bacillus subtilis HPr: use of spectra obtained from mutants to resolve spectral overlap.
复制标题
枯草芽孢杆菌 HPr 的序列特异性 1H NMR 共振分配：使用从突变体获得的光谱来解决光谱重叠问题。
DOI：
10.1021/bi00483a006
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发表时间：
1990
期刊：
影响因子：
2.9
通讯作者：
Klevit,RE
中科院分区：
文献类型：
--
作者：
Wittekind,M;Reizer,J;Klevit,RE
材料和方法蛋白质的纯化和样品制备。通过定点突变产生S46A和S46D突变体，过表达野生型和突变型枯草杆菌Hprs ine。1缩写：PTS，依赖于磷酸烯醇式丙酮酸的糖磷转移酶系统；HPR，PTS中含有组氨酸的磷酸载体蛋白；PEP，磷酸烯醇式丙酮酸；ATP，5‘-三磷酸腺苷；[2H]TSP，3-(三甲基硅基)[2，2，3，3-2H4]丙酸；核磁共振，核磁共振；NOE，核磁共振；2D核磁共振，二维核磁共振光谱；COSY，二维相关光谱；DQF，双量子滤光片；2Q，二维双量子光谱；接力，二维接力相干转移光谱；NOESY，二维NOE光谱；TOCSY，二维全相干光谱。
Materials and MethodsProtein Purification and Sample Preparation. Site-directed mutagenesis to produce the S46A and S46D mutants, overexpression of wild-type and mutant B. subtilis HPrs inE. coli, and their purification have been described (Reizer et al., 1989).1 Abbreviations: PTS, phosphoenolpyruvate-dependent sugarphosphotransferase system; HPr, histidine-containing phosphocarrier protein of the PTS; PEP, phosphoenolpyruvate; ATP, adenosine 5'-triphosphate;[2H] TSP, sodium 3-(trimethylsilyl)[2, 2, 3, 3-2H4] propionate; NMR, nu-clear magnetic resonance; NOE, nuclearOverhauser effect; 2D NMR, two-dimensional nuclear magnetic resonance spectroscopy; COSY, twodimensional./-correlated spectroscopy; DQF, double quantum filter; 2Q, two-dimensional double quantum spectroscopy; RELAY, two-dimen-sional relayed coherence transfer spectroscopy; NOESY, two-dimensional NOE spectroscopy; TOCSY, two-dimensional total coherence spectroscopy.