High-resolution structure of the phosphorylated form of the histidine-containing phosphocarrier protein HPr from Escherichia coli determined by restrained molecular dynamics from NMR-NOE data.
High-resolution structure of the phosphorylated form of the histidine-containing phosphocarrier protein HPr from Escherichia coli determined by restrained molecular dynamics from NMR-NOE data.
复制标题
通过 NMR-NOE 数据的限制分子动力学确定大肠杆菌含组氨酸磷酸化载体蛋白 HPr 的磷酸化形式的高分辨率结构。
DOI：
10.1006/jmbi.1994.0075
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发表时间：
1995
影响因子：
5.6
通讯作者：
G. Robillard
中科院分区：
文献类型：
--
作者：
Nico A. J. van Nuland;R. Boelens;R. Scheek;G. Robillard
含组氨酸的磷酸载体蛋白，HPr，从大肠杆菌的磷酸化形式的解决方案的结构已确定由NMR结合约束的分子动力学模拟。磷-HPr（P-HPr）的结构的结果从分子动力学模拟在水中，使用时间依赖性的距离限制，以达到与测量的NOE协议。从三维1H-1H-15 N HSQC-NOESY和二维1H-1HNOESY光谱中鉴定了实验抑制，并与未磷酸化形式的抑制进行了比较。HPr磷酸化后的结构变化仅限于活性位点，如化学位移、3 JNHH α-偶联常数和NOE模式的变化所证明。化学位移的变化，主要是为质子的位置接近磷酰基连接到His 15咪唑环。可以检测到的差异的NOE的强度，涉及侧链质子的His 15和Pro 18，导致在两个环的相对位置的变化。此外，一个小的变化，可以检测到在三键J-耦合之间的酰胺质子和H α质子的Thr 16和Arg 17磷酸化后，在协议的变化，这两个残基的phi扭转角从时间平均的限制在水中的分子动力学模拟。这些结果不支持在残基16处的扭转角菌株在粪链球菌HPr的机制中的拟议作用。相反，磷酸化似乎在残基His 15处引入扭转角应变。该菌株可以促进磷酰基转移到酶II的A结构域。磷酸-组氨酸不稳定的氢键的侧链基团的Arg 17;相反，稳定的氢键之间形成的磷酸基团和骨干酰胺质子的Thr 16和Arg 17，这表明最大的变化，在磷酸化后的化学位移，和氢键涉及侧链O γ质子的Thr 16。HPr接受来自酶I的磷酰基，并随后将其捐赠给各种酶II物种的A结构域。HPr上EI的结合位点类似于甘露醇特异性酶II的A结构域，这可以从通过异分子单量子相干光谱观察到的酰胺质子和氮化学位移的变化中得出结论。
The solution structure of the phosphorylated form of the histidine-containing phosphocarrier protein, HPr, from Escherichia coli has been determined by NMR in combination with restrained molecular dynamics simulations. The structure of phospho-HPr (P-HPr) results from a molecular dynamics simulation in water, using time-dependent distance restraints to attain agreement with the measured NOEs. Experimental restraints were identified from both three-dimensional 1H-1H-15N HSQC-NOESY and two-dimensional 1H-1HNOESY spectra, and compared with those of the unphosphorylated form. Structural changes upon phosphorylation of HPr are limited to the active site, as evidenced by changes in chemical shifts, in 3JNHH alpha-coupling constants and NOE patterns. Chemical shift changes were obtained mainly for protons that were positioned close to the phosphoryl group attached to the His15 imidazole ring. Differences could be detected in the intensity of the NOEs involving the side-chain protons of His15 and Pro18, resulting from a change in the relative position of the two rings. In addition, a small change could be detected in the three-bond J-coupling between the amide proton and the H alpha proton of Thr16 and Arg17 upon phosphorylation, in agreement with the changes of the phi torsion angle of these two residues obtained from time-averaged restrained molecular dynamics simulations in water. The proposed role of the torsion-angle strain at residue 16 in the mechanism of Streptococcus faecalis HPr is not supported by these results. In contrast, phosphorylation seems to introduce torsion angle strain at residue His15. This strain could facilitate the transfer of the phosphoryl group to the A-domain at enzyme II. The phospho-histidine is not stabilised by hydrogen bonds to the side-chain group of Arg17; instead stable hydrogen bonds are formed between the phosphate group and the backbone amide protons of Thr16 and Arg17, which show the largest changes in chemical shift upon phosphorylation, and a hydrogen bond involving the side-chain O gamma proton of Thr16. HPr accepts the phosphoryl group from enzyme I and donates it subsequently to the A domain of various enzyme II species. The binding site for EI on HPr resembles that of the A domain of the mannitol-specific enzyme II, as can be concluded from the changes on the amide proton and nitrogen chemical shifts observed via heteromolecular single-quantum coherence spectroscopy.
大肠杆菌磷酸烯醇丙酮酸：糖磷酸转移酶系统的含组氨酸蛋白 HPr 的活性位点中羧基末端残基的参与。
影响因子：
2.9
作者：
Anderson,JW;Bhanot,P;Georges,F;Klevit,RE;Waygood,EB
通讯作者：
Waygood,EB
通过同核和异核核磁共振波谱重新检查大肠杆菌含组氨酸蛋白质的二级和三级结构。
影响因子：
2.9
作者：
Hammen,PK;Waygood,EB;Klevit,RE
通讯作者：
Klevit,RE
精氨酸 17 残基参与大肠杆菌磷酸烯醇丙酮酸：糖磷酸转移酶系统的含组氨酸蛋白 HPr 的活性位点。
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Anderson,JW;Pullen,K;Georges,F;Klevit,RE;Waygood,EB
通讯作者：
Waygood,EB