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光转化后日本枪乌贼视紫红质的构象变化

Conformation changes of cuttlefish (Euprymna morsei) rhodopsin following photoconversion.

作者信息

Hiraki K, Hamanaka T, Seidou M, Kito Y

机构信息

Department of Biology, Faculty of Science, Osaka University, Japan.

出版信息

Biochim Biophys Acta. 1991 Apr 29;1077(3):355-61. doi: 10.1016/0167-4838(91)90551-a.

Abstract

Cuttlefish (Euprymna morsei) rhodopsin solubilized in lauryl ester of sucrose and its photoproduct, acid metarhodopsin, were examined by small-angle X-ray scattering and chromatofocusing to investigate the conformation changes of visual pigment following photoconversion. From spectroscopic studies, it was found that more than 93% of Euprymna rhodopsin could be converted to meta form under the condition of red light irradiation at neutral pH. Since almost pure acid metarhodopsin solution was prepared without changing the specimen concentration, the small-angle X-ray scattering intensities of both pigment-detergent complexes were directly compared. The radius of gyration increased on going from rhodopsin to acid metarhodopsin by approximately 1.5%. There were also discernible changes in the secondary peak intensities. The distribution function, derived by the Fourier transformation of intensity data, showed a significant change around 55 A. The maximum linear dimension of the rhodopsin-detergent complex was about 95 A and hardly changed after illumination. Intensity at zero angle did not change after illumination, suggesting that the aggregation did not occur. The change of the intensity profile could be due to the conformational change of the pigment-detergent monomers. The pI value of rhodopsin determined by chromatofocusing was 5.32 and that of acid metarhodopsin was 5.06, indicating that a few carboxyl groups are newly dissociated. The shift of the protein mass and the charge redistribution were observed following photoconversion.

摘要

对溶解于蔗糖月桂酸酯中的乌贼(Euprymna morsei)视紫红质及其光产物酸性变视紫红质进行了小角X射线散射和色谱聚焦分析,以研究光转化后视色素的构象变化。通过光谱研究发现,在中性pH条件下红光照射时,超过93%的乌贼视紫红质可转化为变视紫红质形式。由于在不改变样品浓度的情况下制备了几乎纯的酸性变视紫红质溶液,因此可以直接比较两种色素 - 去污剂复合物的小角X射线散射强度。从视紫红质转变为酸性变视紫红质时,回转半径增加了约1.5%。二级峰强度也有明显变化。通过强度数据的傅里叶变换得到的分布函数在约55 Å处显示出显著变化。视紫红质 - 去污剂复合物的最大线性尺寸约为95 Å,光照后几乎没有变化。光照后零角度处的强度没有变化,这表明没有发生聚集。强度分布的变化可能是由于色素 - 去污剂单体的构象变化。通过色谱聚焦测定的视紫红质的pI值为5.32,酸性变视紫红质的pI值为5.06,这表明有一些羧基新解离。光转化后观察到蛋白质质量的移动和电荷重新分布。

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