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首页> 外文期刊>Biochemistry >Direct Observation of the Structural Change of Tyr174 in the Primary Reaction of Sensory Rhodopsin II
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Direct Observation of the Structural Change of Tyr174 in the Primary Reaction of Sensory Rhodopsin II

机译：直接观察感觉视紫红质II初级反应中Tyr174的结构变化。

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Sensory rhodopsin II (SRII) is a negative phototaxisnreceptor containing retinal as its chromophore, which mediates thenavoidance of blue light. The signal transduction is initiated by thenphotoisomerization of the retinal chromophore, resulting in confor-nmational changes of the protein which are transmitted to a transducernprotein. To gain insight into the SRII sensing mechanism, wenemployed time-resolved ultraviolet resonance Raman spectroscopynmonitoring changes in the protein structure in the picosecond timenrange following photoisomerization.We used a 450 nm pump pulse toninitiate the SRII photocycle and two kinds of probe pulses with wavelengths of 225 and 238 nm to detect spectral changes in thentryptophan and tyrosine bands, respectively. The observed spectral changes of the Raman bands are most likely due to tryptophannand tyrosine residues located in the vicinity of the retinal chromophore, i.e., Trp76, Trp171, Tyr51, or Tyr174. The 225 nm UVRRnspectra exhibited bleaching of the intensity for all the tryptophan bands within the instrumental response time, followed by a partialnrecovery with a time constant of 30 ps and no further changes up to 1 ns. In the 238 nm UVRR spectra, a fast recovering componentnwas observed in addition to the 30 ps time constant component. A comparison between the spectra of theWT and Y174Fmutant ofnSRII indicates that Tyr174 changes its structure and/or environment upon chromophore photoisomerization. These data representnthe first real-time observation of the structural change of Tyr174, of which functional importance was pointed out previously.
机译：感觉视紫红质II（SRII）是一种以视网膜为生色团的负性紫杉碱受体，可介导蓝光的产生。然后通过视网膜发色团的光异构化来引发信号转导，从而导致蛋白质的构象变化，并传递至转导蛋白。为了深入了解SRII的感应机制，我们采用了时间分辨的紫外共振拉曼光谱技术来监测光异构化后皮秒时间范围内蛋白质结构的变化。我们使用了450 nm的泵浦脉冲来激发SRII的光周期和两种波长为225的探测脉冲238 nm和238 nm分别检测色氨酸和酪氨酸谱带的光谱变化。观察到的拉曼谱带的光谱变化最可能是由于色氨酸和酪氨酸残基位于视网膜发色团附近，即Trp76，Trp171，Tyr51或Tyr174。 225 nm UVRRn光谱在仪器响应时间内显示了所有色氨酸条带的强度变白，随后以30 ps的时间常数进行部分恢复，直到1 ns都没有进一步的变化。在238 nm UVRR光谱中，除了30 ps时间常数成分外，还观察到了快速恢复的成分。 nSRII的WT和Y174突变体的光谱比较表明，Tyr174在发色团光异构化后改变了其结构和/或环境。这些数据代表了Tyr174的结构变化的首次实时观察，其功能重要性已在前面指出。

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    《Biochemistry》 |2011年第15期|p.3170-3180|共11页
  • 作者

    Misao Mizuno Yuki Sudo Michio Homma and Yasuhisa Mizutani;

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    †Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, Japan‡Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan§PRESTO, Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan;

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