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首页> 外文期刊>Biochemistry >Overexpression in Yeast, Photocycle, and in Vitro Structural Change of an Avian Putative Magnetoreceptor Cryptochrome4
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Overexpression in Yeast, Photocycle, and in Vitro Structural Change of an Avian Putative Magnetoreceptor Cryptochrome4

机译：酵母，光电周期和鸟类推定的磁受体Cryptochrome4的体外结构变化中的过表达。

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Cryptochromes (CRYs) have been found in a wide variety of living organisms and call function a blue light photoreceptors, circadian clock molecules, or magnetoreceptors. Non-mammalian vertebrates have CRY4 in addition to the CRY1 and CRY2 circadian clock con-Toner-its. Though the function of CRY4 is not well understood, chicken CRY4 (cCRY4) may be a magnetoreceptor because of its high level of expression in the retina and light-dependent structural,changes in retinal homogenates. To farther Characterize the photosensitive nature of cCRY4, we developed an expression system using budding yeast and purified cCRY4 at yields of submilligrams of protein per liter with binding of the flavin adenine dinucleotide (FAD) chromophore. Recombinant cCRY4 dissociated from. anti-cCRY4 C1 mAb, which recognizes the C-terminal region of cCRY4, in a light-dependent manner and showed a light-dependent change in its trypsin digestion pattern, suggesting that cCRY4 changes its conformation with light irradiation in the absence of other retinal factors. Combinatorial analyses with UV-visible spectroscopy and immunoprecipitation revealed that there is chromophore reduction in the cCRY4 photocycle and formation of a flavosemiquinone radical intermediate that is likely accompanied by a conformational change in the carboxyl-terminal region. Thus, cCRY4 seems to be an intrinsically photosensitive and photoswitchable molecule and may exemplify a vertebrate model of cryptochrome with possible function as a photosensor and/or magnetoreceptor.
机译：隐色染料（CRYs）已在多种活生物体中发现，并将其称为蓝光感光器，昼夜节律时钟分子或磁感受器。除CRY1和CRY2昼夜节律钟外，非哺乳动物脊椎动物还具有CRY4。尽管对CRY4的功能尚不十分了解，但鸡CRY4（cCRY​​4）可能是一种磁感受器，因为它在视网膜中的高表达水平以及视网膜匀浆中的光依赖性结构变化。为了进一步表征cCRY4的光敏性质，我们开发了一种使用发芽酵母和纯化的cCRY4的表达系统，每升蛋白的亚微克蛋白产量与黄素腺嘌呤二核苷酸（FAD）发色团结合。重组cCRY4从中解离。抗cCRY4 C1 mAb，以光依赖的方式识别cCRY4的C端区域，并在其胰蛋白酶消化模式上显示出光依赖的变化，这表明cCRY4在没有其他视网膜的情况下通过光照射改变了其构象。因素。紫外可见光谱和免疫沉淀的组合分析表明，cCRY4光循环中的发色团减少，并形成了黄酮半醌自由基中间体，这可能伴随着羧基末端区域的构象变化。因此，cCRY4似乎是一种固有的光敏和可光转换的分子，并可以举例说明隐色岩的脊椎动物模型，其可能具有光传感器和/或磁受体的功能。

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