Properties of the dark and signaling states of photoactive yellow protein probed by solution phase hydrogen/deuterium exchange and mass spectrometry
Properties of the dark and signaling states of photoactive yellow protein probed by solution phase hydrogen/deuterium exchange and mass spectrometry
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DOI：
10.1021/bi0608663
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发表时间：
2006-10-03
期刊：
影响因子：
2.9
通讯作者：
Wysocki, Vicki H.
中科院分区：
文献类型：
--
作者：
Cheng, Guilong;Cusanovich, Michael A.;Wysocki, Vicki H.
光活性黄蛋白从暗态到信号态转化的扰动已经通过溶液相氢/氘交换和质谱确定。野生型和M100 A突变体均用于本研究中，其中突变体在稳态光照下提供超过90%的至漂白状态的转化。我们发现，在野生型和突变体的照明扰动，与更灵活的结构在长寿命的信号（I-2 '）状态一致。在野生型的情况下，检测到的构象变化主要在发色团区域周围。对于M100 A突变体，与野生型相比，光和暗之间的H/D交换差异更广泛;不仅发色团周围受到影响，而且还观察到N-末端和中央β-折叠中氘摄取的显著增加。从这项研究中获得的数据和以前的研究结果的基础上，一系列的事件，导致扰动的PYP发色团光异构化的建议。
The perturbations on conversion from the dark state to the signaling state in photoactive yellow protein have been determined by solution-phase hydrogen/deuterium exchange and mass spectrometry. Both the wild type and M100A mutant are used in this study, with the mutant providing over 90% conversion to the bleached state under steady-state illumination. We found perturbations in both the wild type and the mutant on illumination, consistent with a more flexible structure in the long-lived signaling (I-2') state. In the case of the wild type, the conformational changes detected are mainly around the chromophore region. With the M100A mutant, differences in H/D exchange between the light and dark are more extensive as compared to wild type; not only are the chromophore surroundings affected, but significant increases in deuterium uptake in the N-terminus and central beta-sheet are observed as well. On the basis of the data obtained from this study and previous findings, a sequence of events that leads to the perturbation of PYP following chromophore photoisomerization is proposed.