RESONANCE RAMAN-SPECTRA OF BACTERIORHODOPSINS PRIMARY PHOTOPRODUCT - EVIDENCE FOR A DISTORTED 13-CIS RETINAL CHROMOPHORE
RESONANCE RAMAN-SPECTRA OF BACTERIORHODOPSINS PRIMARY PHOTOPRODUCT - EVIDENCE FOR A DISTORTED 13-CIS RETINAL CHROMOPHORE
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DOI：
10.1073/pnas.79.2.403
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发表时间：
1982-01-01
期刊：
影响因子：
--
通讯作者：
MATHIES, R
中科院分区：
文献类型：
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作者：
BRAIMAN, M;MATHIES, R
采用一种新的低温旋转样品技术，获得了细菌视紫红质初级光产物K的共振拉曼光谱。紫膜在77度。K用空间分离的光化（泵浦）和探测激光束照射。514 nm泵浦光产生细菌视紫红质和K.然后将该混合物旋转通过红色（676 nm）探测光束，其选择性地增强来自K的拉曼散射。连续泵浦-探测技术的主要优点是它可以防止泵浦光激发的荧光掩盖红色探测拉曼散射。K在1516、1294、1012、957和811 cm-1处表现出强的拉曼谱线。C15氘代对K指纹线的影响与13-顺式模型化合物中所见的相关性很好，表明K具有13-顺式发色团。在811和957 cm-1处存在异常强的低波数谱线，这归因于氢的面外摆动，表明该蛋白质在反式→反式之后以扭曲的构象保持发色团。顺式异构化。
The resonance Raman spectrum of bacteriorhodopsin''s primary photoproduct K with a novel low-temperature spinning sample technique was obtained. Purple membrane at 77.degree. K is illuminated with spatially separated actinic (pump) and probe laser beams. The 514-nm pump beam produces a photostationary steady-state mixture of bacteriorhodopsin and K. This mixture is then rotated through the red (676 nm) probe beam, which selectively enhances the Raman scattering from K. The essential advantage of the successive pump-and-probe technique is that it prevents the fluorescence excited by the pump beam from masking the red probe Raman scattering. K exhibits strong Raman lines at 1516, 1294, 1012, 957 and 811 cm-1. The effects of C15 deuteration on K fingerprint lines correlate well with those seen in 13-cis model compounds, indicating that K has a 13-cis chromophore. The presence of unusually strong low-wave number lines at 811 and 957 cm-1, attributable to hydrogen out-of-plane wags, indicates that the protein holds the chromophore in a distorted conformation after trans .fwdarw. cis isomerization.