The Aspartyl Replacement of the Active Site Histidine in Histidine-containing Protein, HPr, of the Escherichia coliPhosphoenolpyruvate:Sugar Phosphotransferase System Can Accept and Donate a Phosphoryl Group
The Aspartyl Replacement of the Active Site Histidine in Histidine-containing Protein, HPr, of the Escherichia coliPhosphoenolpyruvate:Sugar Phosphotransferase System Can Accept and Donate a Phosphoryl Group
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大肠杆菌含组氨酸蛋白 HPr 活性位点组氨酸的天冬氨酰取代磷酸烯醇式丙酮酸：糖磷酸转移酶系统可以接受和捐赠磷酰基
DOI：
10.1074/jbc.274.31.21776
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
E. Waygood
中科院分区：
文献类型：
--
作者：
S. Napper;L. Delbaere;E. Waygood
含组氨酸蛋白HPr中的活性位点残基His 15可以被天冬氨酸取代，并且仍然分别作为酶I和酶IIA葡萄糖的磷酸受体和磷酸供体。其他取代，包括半胱氨酸，谷氨酸，丝氨酸，苏氨酸和酪氨酸，未能显示任何活性。与野生型HPr相比，His 15 → AspHPr的I Km增加了10倍，Vmax降低了1000倍。Asp 15的磷酸化导致了一个自发的内部重排，涉及磷酰基和水分子的损失，这是由质谱证实。形成的蛋白质种类具有比His 15 → Asp HPr更高的pI，这可能是由于形成琥珀酰亚胺或异酰亚胺而引起的。水解分离的高pI形式仅在残基15处产生天冬氨酸，未检测到异天冬氨酸。这表明形成了异酰亚胺而不是琥珀酰亚胺。在不存在磷酸化的情况下，没有发现高pI形式的形成，表明磷酸化催化环化的形成。Asn 12参与Asp 15内环化的可能性被His 15 → AspHPr中的Asn 12 → Ala突变所消除。野生型HPr中丙氨酸、天冬氨酸、丝氨酸和苏氨酸的Asn 12取代表明，通常需要能够与His 15的Nε2原子形成氢键的残基，但氢键的消除仅使ink cat/Km降低4倍。
The active site residue, His15, in histidine-containing protein, HPr, can be replaced by aspartate and still act as a phosphoacceptor and phosphodonor with enzyme I and enzyme IIAglucose, respectively. Other substitutions, including cysteine, glutamate, serine, threonine, and tyrosine, failed to show any activity. Enzyme I K m for His15 → Asp HPr is increased 10-fold andV max is decreased 1000-fold compared with wild type HPr. The phosphorylation of Asp15 led to a spontaneous internal rearrangement involving the loss of the phosphoryl group and a water molecule, which was confirmed by mass spectrometry. The protein species formed had a higher pI than His15 → Asp HPr, which could arise from the formation of a succinimide or an isoimide. Hydrolysis of the isolated high pI form gave only aspartic acid at residue 15, and no isoaspartic acid was detected. This indicates that an isoimide rather than a succinimide is formed. In the absence of phosphorylation, no formation of the high pI form could be found, indicating that phosphorylation catalyzed the formation of the cyclization. The possible involvement of Asn12 in an internal cyclization with Asp15 was eliminated by the Asn12 → Ala mutation in His15 → AspHPr. Asn12 substitutions of alanine, aspartate, serine, and threonine in wild type HPr indicated a general requirement for residues capable of forming a hydrogen bond with the Nε2 atom of His15, but elimination of the hydrogen bond has only a 4-fold decrease ink cat/K m .
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L-异天冬氨酸-（D-天冬氨酸）O-甲基转移酶催化自发脱酰胺化 HPr 磷酸载体蛋白的修复。
DOI：
--
发表时间：
1994
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Brennan,TV;Anderson,JW;Jia,Z;Waygood,EB;Clarke,S
通讯作者：
Clarke,S
DOI：
10.3109/10409239109081719
发表时间：
1991
影响因子：
6.5
作者：
H. Wright;D. Urry
通讯作者：
H. Wright;D. Urry
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
DOI：
10.1016/s0021-9258(19)75855-4
发表时间：
1987-01
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
T. Geiger;S. Clarke
通讯作者：
T. Geiger;S. Clarke
通过细菌磷酸转移酶系统进行糖转运。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Weigel,N;Kukuruzinska,MA;Nakazawa,A;Waygood,EB;Roseman,S
通讯作者：
Roseman,S