Sequential backbone resonance assignments of the E.coli dihydrofolate reductase Gly67Val mutant:folate complex
Sequential backbone resonance assignments of the E.coli dihydrofolate reductase Gly67Val mutant:folate complex
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大肠杆菌二氢叶酸还原酶 Gly67Val 突变体的连续主链共振分配：叶酸复合物
DOI：
10.1007/s12104-015-9650-y
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Akasaka,K.
中科院分区：
文献类型：
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作者：
Narayanan,S.P.;Maeno,A.;Wada,Y.;Tate,S.;Akasaka,K.
偶尔，暴露的环区的突变会导致蛋白质功能和/或稳定性的显著变化。E基因的单一突变Gly67Val。暴露的CD环中的秋水仙碱叶酸还原酶(DHFR)就是这样一个例子。我们对E的Gly67Val突变体的HN，NH，Cα和Cβ原子进行了化学位移指认。在pH 7.0、35℃的条件下，将ColiDHFR与叶酸络合，然后评估突变引起的HN、NH、Cα和Cβ化学位移的变化。结果表明，在总体二级结构保持不变的情况下，Gly67Val单突变导致腺苷结合亚区(38-88位残基)周围的多肽骨架发生位点特异性构象变化，而不是在远端催化区域。
Occasionally, a mutation in an exposed loop region causes a significant change in protein function and/or stability. A single mutation Gly67Val ofE. colidihydrofolate reductase (DHFR) in the exposed CD loop is such an example. We have carried out the chemical shift assignments for HN, NH, Cαand Cβatoms of the Gly67Val mutant ofE. coliDHFR complexed with folate at pH 7.0, 35 °C, and then evaluated the HN, NH, Cαand Cβchemical shift changes caused by the mutation. The result indicates that, while the overall secondary structure remains the same, the single mutation Gly67Val causes site-specific conformational changes of the polypeptide backbone restricted around the adenosine-binding subdomain (residues 38–88) and not in the distant catalytic domain.