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1
Aspartic acids 96 and 85 play a central role in the function of bacteriorhodopsin as a proton pump.天冬氨酸96和85在细菌视紫红质作为质子泵的功能中起核心作用。
EMBO J. 1989 Jun;8(6):1657-63. doi: 10.1002/j.1460-2075.1989.tb03556.x.
2
Substitution of amino acids Asp-85, Asp-212, and Arg-82 in bacteriorhodopsin affects the proton release phase of the pump and the pK of the Schiff base.细菌视紫红质中氨基酸天冬氨酸-85、天冬氨酸-212和精氨酸-82的取代会影响泵的质子释放阶段以及席夫碱的pK值。
Proc Natl Acad Sci U S A. 1990 Feb;87(3):1018-22. doi: 10.1073/pnas.87.3.1018.
3
Properties of Asp212----Asn bacteriorhodopsin suggest that Asp212 and Asp85 both participate in a counterion and proton acceptor complex near the Schiff base.天冬氨酸212突变为天冬酰胺的细菌视紫红质的特性表明，天冬氨酸212和天冬氨酸85都参与了席夫碱附近的抗衡离子和质子受体复合物。
J Biol Chem. 1991 Jun 25;266(18):11478-84.
4
A defective proton pump, point-mutated bacteriorhodopsin Asp96----Asn is fully reactivated by azide.一种存在点突变的缺陷型质子泵——细菌视紫红质Asp96突变为Asn，可被叠氮化物完全重新激活。
EMBO J. 1989 Nov;8(11):3477-82. doi: 10.1002/j.1460-2075.1989.tb08512.x.
5
Connectivity of the retinal Schiff base to Asp85 and Asp96 during the bacteriorhodopsin photocycle: the local-access model.细菌视紫红质光循环过程中视网膜席夫碱与Asp85和Asp96的连接性：局部可及模型
Biophys J. 1998 Sep;75(3):1455-65. doi: 10.1016/S0006-3495(98)74064-0.
6
Aspartic acid-96 is the internal proton donor in the reprotonation of the Schiff base of bacteriorhodopsin.天冬氨酸-96是细菌视紫红质席夫碱再质子化过程中的内部质子供体。
Proc Natl Acad Sci U S A. 1989 Dec;86(23):9228-32. doi: 10.1073/pnas.86.23.9228.
7
Asp 46 can substitute Asp 96 as the Schiff base proton donor in bacteriorhodopsin.在细菌视紫红质中，天冬氨酸46可以替代天冬氨酸96作为席夫碱质子供体。
Biochemistry. 1995 Nov 28;34(47):15599-606. doi: 10.1021/bi00047a027.
8
Protein changes associated with reprotonation of the Schiff base in the photocycle of Asp96-->Asn bacteriorhodopsin. The MN intermediate with unprotonated Schiff base but N-like protein structure.与天冬氨酸96→天冬酰胺细菌视紫红质光循环中席夫碱再质子化相关的蛋白质变化。具有未质子化席夫碱但N样蛋白质结构的MN中间体。
J Biol Chem. 1992 Oct 15;267(29):20782-6.
9
Aspartic acid substitutions affect proton translocation by bacteriorhodopsin.天冬氨酸替代影响细菌视紫红质的质子转运。
Proc Natl Acad Sci U S A. 1988 Jun;85(12):4148-52. doi: 10.1073/pnas.85.12.4148.
10
Replacement of aspartic acid-96 by asparagine in bacteriorhodopsin slows both the decay of the M intermediate and the associated proton movement.细菌视紫红质中天冬氨酸-96被天冬酰胺取代会减缓M中间体的衰减以及相关的质子移动。
Proc Natl Acad Sci U S A. 1989 Apr;86(7):2167-71. doi: 10.1073/pnas.86.7.2167.

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1
Engineering of soluble bacteriorhodopsin.可溶性细菌视紫红质的工程改造
Chem Sci. 2025 May 13. doi: 10.1039/d5sc02453f.
2
Excited-state dynamics of all-trans protonated retinal Schiff base in CRABPII-based rhodopsin mimics.全反式质子化视黄醛 Schiff 碱在 CRABPII 模拟视紫红质中的激发态动力学。
Biophys J. 2022 Nov 1;121(21):4109-4118. doi: 10.1016/j.bpj.2022.09.032. Epub 2022 Sep 30.
3
The Voltage Dependent Sidedness of the Reprotonation of the Retinal Schiff Base Determines the Unique Inward Pumping of Xenorhodopsin.视黄醛 Schiff 碱的返质子化的电压依赖性侧决定了 Xenorhodopsin 的独特内向泵浦。
Angew Chem Int Ed Engl. 2021 Oct 11;60(42):23010-23017. doi: 10.1002/anie.202103882. Epub 2021 Sep 15.
4
Color-tuning of natural variants of heliorhodopsin.天然视紫红质变体的颜色调谐。
Sci Rep. 2021 Jan 13;11(1):854. doi: 10.1038/s41598-020-72125-0.
5
Diversity, Mechanism, and Optogenetic Application of Light-Driven Ion Pump Rhodopsins.光驱动离子泵视紫红质的多样性、机制及光遗传学应用。
Adv Exp Med Biol. 2021;1293:89-126. doi: 10.1007/978-981-15-8763-4_6.
6
Isotope-Labeled Aspartate Sidechain as a Non-Perturbing Infrared Probe: Application to Investigate the Dynamics of a Carboxylate Buried Inside a Protein.同位素标记的天冬氨酸侧链作为一种非干扰性红外探针：用于研究埋藏在蛋白质内部的羧酸盐动力学的应用。
Chem Phys Lett. 2017 Sep 1;683:193-198. doi: 10.1016/j.cplett.2017.03.064. Epub 2017 Mar 23.
7
Schiff Base Proton Acceptor Assists Photoisomerization of Retinal Chromophores in Bacteriorhodopsin.席夫碱质子受体协助细菌视紫红质中视黄醛发色团的光异构化。
Biophys J. 2017 Jun 20;112(12):2503-2519. doi: 10.1016/j.bpj.2017.05.015.
8
Conversion of a light-driven proton pump into a light-gated ion channel.将光驱动质子泵转化为光门控离子通道。
Sci Rep. 2015 Nov 24;5:16450. doi: 10.1038/srep16450.
9
Recent advances in engineering microbial rhodopsins for optogenetics.用于光遗传学的工程化微生物视紫红质的最新进展。
Curr Opin Struct Biol. 2015 Aug;33:8-15. doi: 10.1016/j.sbi.2015.05.001. Epub 2015 Jun 1.
10
Microbial and animal rhodopsins: structures, functions, and molecular mechanisms.微生物和动物视紫红质：结构、功能及分子机制
Chem Rev. 2014 Jan 8;114(1):126-63. doi: 10.1021/cr4003769. Epub 2013 Dec 23.

本文引用的文献

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The halo-opsin gene. II. Sequence, primary structure of halorhodopsin and comparison with bacteriorhodopsin.嗜盐菌视紫红质基因。II. 嗜盐菌视紫红质的序列、一级结构及其与细菌视紫红质的比较。
EMBO J. 1987 Jan;6(1):265-73. doi: 10.1002/j.1460-2075.1987.tb04749.x.
2
Fast stages of photoelectric processes in biological membranes. I. Bacteriorhodopsin.生物膜中光电过程的快速阶段。I. 细菌视紫红质
Eur J Biochem. 1981 Jul;117(3):461-70. doi: 10.1111/j.1432-1033.1981.tb06361.x.
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Photocurrents induced on black lipid membranes by purple membranes: a method of reconstitution and a kinetic study of the photocurrents.
Ann N Y Acad Sci. 1980;358:324-7. doi: 10.1111/j.1749-6632.1980.tb15405.x.
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Reversible photolysis of the purple complex in the purple membrane of Halobacterium halobium.嗜盐菌紫色膜中紫色复合物的可逆光解作用。
Eur J Biochem. 1973 Aug 17;37(2):316-26. doi: 10.1111/j.1432-1033.1973.tb02990.x.
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Functions of a new photoreceptor membrane.一种新型光感受器膜的功能。
Proc Natl Acad Sci U S A. 1973 Oct;70(10):2853-7. doi: 10.1073/pnas.70.10.2853.
6
Isolation of the cell membrane of Halobacterium halobium and its fractionation into red and purple membrane.嗜盐菌细胞膜的分离及其分成红色膜和紫色膜的分级分离。
Methods Enzymol. 1974;31:667-78. doi: 10.1016/0076-6879(74)31072-5.
7
Direct measurement of electric current generation by cytochrome oxidase, H+-ATPase and bacteriorhodopsin.通过细胞色素氧化酶、H⁺-ATP酶和细菌视紫红质对电流产生的直接测量。
Nature. 1974 May 24;249(455):321-4. doi: 10.1038/249321a0.
8
Time-resolved photoelectric and absorption signals from oriented purple membranes immobilized in gel.来自固定在凝胶中的定向紫膜的时间分辨光电信号和吸收信号。
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9
Light-driven protonation changes of internal aspartic acids of bacteriorhodopsin: an investigation by static and time-resolved infrared difference spectroscopy using [4-13C]aspartic acid labeled purple membrane.细菌视紫红质内部天冬氨酸的光驱动质子化变化：使用[4-13C]天冬氨酸标记的紫膜通过静态和时间分辨红外差光谱进行的研究
Biochemistry. 1985 Jan 15;24(2):400-7. doi: 10.1021/bi00323a024.
10
Aspartic acid substitutions affect proton translocation by bacteriorhodopsin.天冬氨酸替代影响细菌视紫红质的质子转运。
Proc Natl Acad Sci U S A. 1988 Jun;85(12):4148-52. doi: 10.1073/pnas.85.12.4148.

天冬氨酸96和85在细菌视紫红质作为质子泵的功能中起核心作用。

Aspartic acids 96 and 85 play a central role in the function of bacteriorhodopsin as a proton pump.

作者信息

Butt H J, Fendler K, Bamberg E, Tittor J, Oesterhelt D

机构信息

Max-Plank-Institut für Biophysik, Frankfurt am Main, FRG.

出版信息

EMBO J. 1989 Jun;8(6):1657-63. doi: 10.1002/j.1460-2075.1989.tb03556.x.

DOI:10.1002/j.1460-2075.1989.tb03556.x
PMID:2548851
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC401006/
Abstract

A spectroscopic and functional analysis of two point-mutated bacteriorhodopsins (BRs) from phototrophic negative halobacterial strains is reported. Bacteriorhodopsin from strain 384 contains a glutamic acid instead of an aspartic acid at position 85 and BR from strain 326 contains asparagine instead of aspartic acid at position 96. Compared to wild-type BR, the M formation in BR Asp85---Glu is accwelerated approximately 10-fold, whereas the M decay in BR Asp96---Asn is slowed down approximately 50-fold at pH6. Purple membrane sheets containing the mutated BRs were oriented and immobilized in polyacrylamide gels or adsorbed to planar lipid films. The measured kinetics of the photocurrents under various conditions agree with the observed photocycle kinetics. The ineffectivity of BR Asp85---Glu resides in the dominance of an inactive species absorbing maximally at approximately 610 nm, while BR Asp96---Asn is ineffective due to its slow photocycle. These experimental results suggest that aspartic acid 96 plays a crucial role for the reprotonation of the Schiff base. Both residues are essential for an effective proton pump.

摘要

报道了对来自光合阴性嗜盐菌菌株的两种点突变细菌视紫红质（BRs）的光谱和功能分析。384菌株的细菌视紫红质在85位含有谷氨酸而非天冬氨酸，326菌株的BR在96位含有天冬酰胺而非天冬氨酸。与野生型BR相比，BR Asp85---Glu中M的形成加速了约10倍，而在pH6时，BR Asp96---Asn中M的衰减减慢了约50倍。含有突变BRs的紫膜片在聚丙烯酰胺凝胶中定向固定或吸附到平面脂质膜上。在各种条件下测量的光电流动力学与观察到的光循环动力学一致。BR Asp85---Glu的无效性在于一种在约610nm处吸收最大的无活性物种占主导地位，而BR Asp96---Asn由于其缓慢的光循环而无效。这些实验结果表明，天冬氨酸96对席夫碱的再质子化起着关键作用。这两个残基对于有效的质子泵都是必不可少的。