荚膜红杆菌细胞色素bc1喹啉氧化位点质子转移氢键网络的鉴定。

The Journal of Biological Chemistry Pub Date : 2023-10-01 Epub Date: 2023-09-14 DOI:10.1016/j.jbc.2023.105249
Arkadiusz Borek, Anna Wójcik-Augustyn, Patryk Kuleta, Robert Ekiert, Artur Osyczka
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引用次数: 0

摘要

细胞色素bc1催化从醌(QH2)到细胞色素c的电子转移,这些反应与穿过节能膜的质子易位相结合。催化循环的能量效率是由两个电子和两个质子的分岔反应保证的,该分岔反应导致QH2的氧化和Rieske簇和血红素bL的还原。与该反应相关的质子路径仍然难以捉摸。在这里,我们使用定点诱变和量子力学计算来分析胶囊红杆菌细胞色素bc1中位于QH2氧化位点的血红素bL侧的可质子化侧链的贡献。我们观察到,当H276和E295同时突变为H276F/E295V双突变体中的不可调性残基时,质子路径被有效地切断。两个单一突变体H276F或E295V效率较低,但仍以功能相关的速率转移质子。自然选择暴露了两个单一突变,N279S和M154T,它们恢复了H276F/E295V中的功能性质子转移。量子力学计算表明,H276F/E295V将Y147的侧链捕获在远离QH2的位置,而N279S或M154T诱导局部变化,将Y147从该位置释放。这缩短了Y147和D278的可质子化基团之间的距离和/或增加了Y147侧链的迁移率,这使得Y147在H276F/E295V中有效地将质子从QH2转移到D278。总体而言,我们的研究确定了由E295、H276、D278和Y147建立的扩展氢键网络,该网络参与在QH2氧化位点的血红素bL侧从QH2有效去除质子。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Identification of hydrogen bonding network for proton transfer at the quinol oxidation site of Rhodobacter capsulatus cytochrome bc<sub>1</sub>.

Identification of hydrogen bonding network for proton transfer at the quinol oxidation site of Rhodobacter capsulatus cytochrome bc<sub>1</sub>.

Identification of hydrogen bonding network for proton transfer at the quinol oxidation site of Rhodobacter capsulatus cytochrome bc<sub>1</sub>.

Identification of hydrogen bonding network for proton transfer at the quinol oxidation site of Rhodobacter capsulatus cytochrome bc1.

Cytochrome bc1 catalyzes electron transfer from quinol (QH2) to cytochrome c in reactions coupled to proton translocation across the energy-conserving membrane. Energetic efficiency of the catalytic cycle is secured by a two-electron and two-proton bifurcation reaction leading to oxidation of QH2 and reduction of the Rieske cluster and heme bL. The proton paths associated with this reaction remain elusive. Here, we used site-directed mutagenesis and quantum mechanical calculations to analyze the contribution of protonable side chains located at the heme bL side of the QH2 oxidation site in Rhodobacter capsulatus cytochrome bc1. We observe that the proton path is effectively switched off when H276 and E295 are simultaneously mutated to the nonprotonable residues in the H276F/E295V double mutant. The two single mutants, H276F or E295V, are less efficient but still transfer protons at functionally relevant rates. Natural selection exposed two single mutations, N279S and M154T, that restored the functional proton transfers in H276F/E295V. Quantum mechanical calculations indicated that H276F/E295V traps the side chain of Y147 in a position distant from QH2, whereas either N279S or M154T induce local changes releasing Y147 from that position. This shortens the distance between the protonable groups of Y147 and D278 and/or increases mobility of the Y147 side chain, which makes Y147 efficient in transferring protons from QH2 toward D278 in H276F/E295V. Overall, our study identified an extended hydrogen bonding network, build up by E295, H276, D278, and Y147, involved in efficient proton removal from QH2 at the heme bL side of QH2 oxidation site.

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