NMR studies of the Escherichia coli trp aporepressor. Sequence-specific assignment of the aromatic proton resonances.
NMR studies of the Escherichia coli trp aporepressor. Sequence-specific assignment of the aromatic proton resonances.
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大肠杆菌 trp 阻压剂的 NMR 研究。
DOI：
10.1111/j.1432-1033.1989.tb21083.x
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发表时间：
1989
期刊：
影响因子：
--
通讯作者：
Jardetzky,O
中科院分区：
文献类型：
--
作者：
Hyde,EI;Ramesh,V;Roberts,GC;Arrowsmith,CH;Treat-Clemons,L;Klaic,B;Jardetzky,O
通过二维相关光谱（COSY）、home-Hartmann-Hahn（HOHAHA）光谱和核Overhauser增强光谱（NOESY）技术以及化学位移的pH依赖性研究，结合蛋白质的选择性氘代，大肠杆菌trpaporepressor的1H-NMR光谱的芳香区中的共振被指定为氨基酸类型。通过将观察到的残基间NOE与基于蛋白质晶体结构预期的NOE进行比较，完成了芳香族共振的完整序列特异性分配[Zhang，R. -G.，Joachimiak，A.，劳森角L.，舍维茨河W.，Otwinowski，Z. & Sigler，P. B.（1987）Nature 327，591-597]。后者的实验还允许对一些高场甲基共振进行序列特异性分配。芳香族区域的光谱，特别是从二聚体界面的残基的共振的完整分配，开辟了详细的研究辅阻遏物和运营商结合的构象效应的方式。
The resonances in the aromatic region of the1H‐NMR spectrum of theEscherichia coli trpaporepressor have been assigned to amino acid type by two‐dimensional correlated spectroscopy (COSY), homonuclear Hartmann‐Hahn (HOHAHA) spectroscopy and nuclear Overhauser enhancement spectroscopy (NOESY) techniques and studies of the pH dependence of the chemical shifts, in combination with selective deuteration of the protein. Complete sequence‐specific assignments of the aromatic resonances have been made by comparing the observed inter‐residue NOEs with those expected on the basis of the crystal structure of the protein [Zhang, R.‐G., Joachimiak, A., Lawson, C. L., Shevitz, R. W., Otwinowski, Z. & Sigler, P. B. (1987)Nature 327, 591–597]. The latter experiments have also permitted the sequence‐specific assignment of some of the high‐field methyl resonances. The complete assignment of the aromatic region of the spectrum, in particular of resonances from residues at the dimer interface, opens the way to detailed studies of the conformational effects of corepressor and operator binding.