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Evidence for light-induced 13-cis, 14-s-cis isomerization in bacteriorhodopsin obtained by FTIR difference spectroscopy using isotopically labelled retinals.利用同位素标记的视黄醛的 FTIR 差谱法获得的细菌视紫红质中光诱导的 13-cis、14-s-cis 异构化的证据。
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The back photoreaction of the M intermediate in the photocycle of bacteriorhodopsin: mechanism and evidence for two M species.细菌视紫红质光循环中M中间体的反向光反应：两种M物种的机制与证据
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Closing in on bacteriorhodopsin: progress in understanding the molecule.走近细菌视紫红质：对该分子认识的进展
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Azide reduces the hydrophobic barrier of the bacteriorhodopsin proton channel.叠氮化物降低了细菌视紫红质子通道的疏水屏障。
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Site-directed spin-labeling reveals the orientation of the amino acid side-chains in the E-F loop of bacteriorhodopsin.定点自旋标记揭示了细菌视紫红质E-F环中氨基酸侧链的取向。
J Mol Biol. 1999 Mar 19;287(1):163-71. doi: 10.1006/jmbi.1998.2593.
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Protein conformational changes in the bacteriorhodopsin photocycle.细菌视紫红质光循环中的蛋白质构象变化。
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10
Conformational change of helix G in the bacteriorhodopsin photocycle: investigation with heavy atom labeling and x-ray diffraction.细菌视紫红质光循环中螺旋G的构象变化：重原子标记与X射线衍射研究
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通过时间分辨电子顺磁共振光谱法揭示细菌视紫红质的光激发构象变化。

Unraveling photoexcited conformational changes of bacteriorhodopsin by time resolved electron paramagnetic resonance spectroscopy.

作者信息

Rink T, Pfeiffer M, Oesterhelt D, Gerwert K, Steinhoff H J

机构信息

Lehrstuhl für Biophysik, Ruhr-Universität Bochum, 44801 Bochum, Germany.

出版信息

Biophys J. 2000 Mar;78(3):1519-30. doi: 10.1016/S0006-3495(00)76704-X.

DOI:10.1016/S0006-3495(00)76704-X
PMID:10692336
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1300749/
Abstract

By means of time-resolved electron paramagnetic resonance (EPR) spectroscopy, the photoexcited structural changes of site-directed spin-labeled bacteriorhodopsin are studied. A complete set of cysteine mutants of the C-D loop, positions 100-107, and of the E-F loop, including the first alpha-helical turns of helices E and F, positions 154-171, was modified with a methanethiosulfonate spin label. The EPR spectral changes occurring during the photocycle are consistent with a small movement of helix C and an outward tilt of helix F. These helix movements are accompanied by a rearrangement of the E-F loop and of the C-terminal turn of helix E. The kinetic analysis of the transient EPR data and the absorbance changes in the visible spectrum reveals that the conformational change occurs during the lifetime of the M intermediate. Prominent rearrangements of nitroxide side chains in the vicinity of D96 may indicate the preparation of the reprotonation of the Schiff base. All structural changes reverse with the recovery of the bacteriorhodopsin initial state.

摘要

通过时间分辨电子顺磁共振（EPR）光谱法，研究了定点自旋标记细菌视紫红质的光激发结构变化。用甲硫基磺酸盐自旋标记修饰了C-D环（第100-107位）和E-F环（包括螺旋E和F的第一个α-螺旋转角，第154-171位）的一组完整的半胱氨酸突变体。光循环过程中发生的EPR光谱变化与螺旋C的微小移动和螺旋F的向外倾斜一致。这些螺旋运动伴随着E-F环和螺旋E的C末端转角的重排。对瞬态EPR数据和可见光谱中吸光度变化的动力学分析表明，构象变化发生在M中间体的寿命期间。D96附近的氮氧化物侧链的显著重排可能表明席夫碱再质子化的准备过程。所有结构变化都随着细菌视紫红质初始状态的恢复而逆转。