Fourier transform infrared difference spectroscopy of bacteriorhodopsin and its photoproducts regenerated with deuterated tyrosine.
Fourier transform infrared difference spectroscopy of bacteriorhodopsin and its photoproducts regenerated with deuterated tyrosine.
复制标题
细菌视紫红质及其用氘代酪氨酸再生的光产物的傅里叶变换红外差异光谱。
DOI：
10.1021/bi00369a028
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Termini,J
中科院分区：
文献类型：
--
作者：
Dollinger,G;Eisenstein,L;Lin,SL;Nakanishi,K;Termini,J
哥伦比亚大学化学系，纽约，纽约 10027 1986 年 1 月 13 日收稿；修订稿于 1986 年 5 月 28 日收到摘要：傅里叶变换红外 (FTIR) 差值光谱已被用于检测由于蛋白质中的酪氨酸残基引起的振动模式，这些残基在光适应细菌视紫红质 (LA) 和其他物种（即 K 和 M 中间体以及暗适应细菌视紫红质 (DA)）之间位置或强度发生变化。为了帮助识别这些不同物种中变化的谱带，在低和高 pH 值的 H20 和 D20 中测量了游离氨基酸 Tyr-í/0、Tyr-d2（OH 邻位 2H）和 Tyr-dA（OH 邻位和间位 2H）的 FTIR 光谱。然后使用以这种方式获得的酪氨酸种类的特征频率来识别各种细菌视紫红质种类中酪氨酸残基的质子化状态的变化。两个诊断上最有用的条带是 Tyr (OH)-¿ 2 的 ~ 1480-cm-1 条带和 Tyr (O~)-Bacteriorhodopsin (BR) 1 的~ 1277-cm-1 条带，它是来自盐杆菌紫色膜中包含的唯一蛋白质。它一直是深入研究的焦点，主要是因为它起到光激活质子泵的作用，这是在视网膜异构化时发生的事件。质子泵浦产生的电位差为 ATP 的合成提供能量。结合 BR 蛋白和 ATP 酶的合成囊泡在 ADP 和无机磷酸盐存在下在光照下产生 ATP，从而支持化学分析假说
Department of Chemistry, Columbia University, New York, New York 10027 Received January 13, 1986; Revised Manuscript Received May 28, 1986 abstract: Fourier transform infrared (FTIR) difference spectroscopyhas been qsed to detect the vibrational modes due to tyrosine residues in the protein that change in position or intensity between light-adapted bacteriorhodopsin (LA) and other species, namely, the K and M intermediates and dark-adapted bacteriorhodopsin (DA). To aid in the identification of the bands that change in these various species, the FTIR spectra of the free amino acids Tyr-í/0, Tyr-d2 (2H at positions ortho to OH), and Tyr-dA (2H at positions ortho and meta to OH) were measured in H20 and D20 at low and high pH. The characteristicfrequencies of the Tyr species obtained in this manner were then used to identify the changes in protonation state of the tyrosine residues in the various bacteriorhodopsin species. The two diagnostically most useful bands were the~ 1480-cm-1 band of Tyr (OH)-¿ 2 and the~ 1277-cm_1 band of Tyr (O~)-Bacteriorhodopsin (BR) 1 is the sole protein contained in the purple membrane from the bacteria Halobacterium halobium. It has been the focus of intense investigation primarily because it functions as a light-activated proton pump, an event that takes place upon the isomerization of retinal. The potential difference that occurs as a result of proton pumping provides the energy for the synthesis of ATP. Synthetic vesicles in-corporating BR protein along with ATPase are seen to produce ATP in thepresence of ADP and inorganic phosphate upon illumination, thus supporting the chemiosmptic hypothesis of