当前位置:
X-MOL 学术
›
Biochemistry
›
论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Loss of Fourth Electron-Transferring Tryptophan in Animal (6–4) Photolyase Impairs DNA Repair Activity in Bacterial Cells
Biochemistry ( IF 2.9 ) Pub Date : 2017-09-20 00:00:00 , DOI: 10.1021/acs.biochem.7b00366 Junpei Yamamoto 1 , Kohei Shimizu 1 , Takahiro Kanda 1 , Yuhei Hosokawa 1 , Shigenori Iwai 1 , Pascal Plaza 2, 3 , Pavel Müller 4
Biochemistry ( IF 2.9 ) Pub Date : 2017-09-20 00:00:00 , DOI: 10.1021/acs.biochem.7b00366 Junpei Yamamoto 1 , Kohei Shimizu 1 , Takahiro Kanda 1 , Yuhei Hosokawa 1 , Shigenori Iwai 1 , Pascal Plaza 2, 3 , Pavel Müller 4
Affiliation
(6–4) photolyases [(6–4)PLs] are flavoproteins that use blue light to repair the ultraviolet-induced pyrimidine(6–4)pyrimidone photoproduct in DNA. Their flavin adenine dinucleotide (FAD) cofactor can be reduced to its repair-active FADH– form by a photoinduced electron transfer reaction. In animal (6–4)PLs, a chain of four Trp residues was suggested to be involved in a stepwise transfer of an oxidation hole from the flavin to the surface of the protein. Here, we investigated the effect of mutation of the fourth Trp on the DNA photorepair activity of Xenopus laevis (6–4)PL (Xl64) in bacterial cells. The photoreduction and photorepair properties of this mutant protein were independently characterized in vitro. Our results demonstrate that the mutation of the fourth Trp in Xl64 drastically impairs the DNA repair activity in cells and that this effect is due to the inhibition of the photoreduction process. We thereby show that the photoreductive formation of FADH– through the Trp tetrad is essential for the biological function of the animal (6–4)PL. The role of the Trp cascade, and of the fourth Trp in particular, is discussed.
中文翻译:
动物(6–4)光裂解酶中第四电子转移色氨酸的损失会损害细菌细胞中的DNA修复活性。
(6–4)光解酶[(6–4)PLs]是黄蛋白,使用蓝光修复DNA中紫外线诱导的嘧啶(6–4)嘧啶酮光产物。他们的黄素腺嘌呤二核苷酸(FAD)辅因子可以被减少到其修复活性FADH -通过光诱导电子转移反应形式。在动物(6–4)PL中,建议将四个Trp残基链与黄素到蛋白质表面的氧化孔逐步转移有关。在这里,我们研究了第四种Trp突变对非洲爪蟾(6–4)PL(Xl 64)DNA光修复活性的影响。该突变蛋白的光还原和光修复特性在体外被独立地表征。我们的结果证明,Xl 64中第四个Trp的突变极大地损害了细胞中的DNA修复活性,并且这种作用是由于抑制了光还原过程。我们由此表明,FADH的光还原形成-通过色氨酸四分体是动物(6-4)PL的生物学功能是必不可少的。讨论了Trp级联的作用,尤其是第四个Trp的作用。
更新日期:2017-09-20
中文翻译:
动物(6–4)光裂解酶中第四电子转移色氨酸的损失会损害细菌细胞中的DNA修复活性。
(6–4)光解酶[(6–4)PLs]是黄蛋白,使用蓝光修复DNA中紫外线诱导的嘧啶(6–4)嘧啶酮光产物。他们的黄素腺嘌呤二核苷酸(FAD)辅因子可以被减少到其修复活性FADH -通过光诱导电子转移反应形式。在动物(6–4)PL中,建议将四个Trp残基链与黄素到蛋白质表面的氧化孔逐步转移有关。在这里,我们研究了第四种Trp突变对非洲爪蟾(6–4)PL(Xl 64)DNA光修复活性的影响。该突变蛋白的光还原和光修复特性在体外被独立地表征。我们的结果证明,Xl 64中第四个Trp的突变极大地损害了细胞中的DNA修复活性,并且这种作用是由于抑制了光还原过程。我们由此表明,FADH的光还原形成-通过色氨酸四分体是动物(6-4)PL的生物学功能是必不可少的。讨论了Trp级联的作用,尤其是第四个Trp的作用。