Cleavage of the retinal pigment epithelium-specific protein RPE65 under oxidative stress.
Cleavage of the retinal pigment epithelium-specific protein RPE65 under oxidative stress.
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DOI：
10.1016/j.ijbiomac.2010.05.014
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发表时间：
2010-08-01
影响因子：
8.2
通讯作者：
Jahng WJ
中科院分区：
文献类型：
--
作者：
Lee H;Chung H;Arnouk H;Lamoke F;Hunt RC;Hrushesky WJ;Wood PA;Lee SH;Jahng WJ
视紫红质11-顺式视黄酰基亚胺发色团在视觉周期中的再生及其控制机制是视觉研究领域的核心问题。视网膜色素上皮（RPE）特异性蛋白RPE65主要参与所有反式视黄醇酯的异构化和水解。在这项研究中，我们研究了氧化应激条件下RPE65的裂解及其潜在的调控机制。将D407 RPE细胞培养液暴露于H2O2（100-1000µM）中。采用凝胶电泳和western blotting检测RPE65及凋亡相关蛋白水平的变化。质谱分析证实了RPE65的性质。采用C57BL/6J （M450）和C3HeB/FeJ （L450）小鼠进行体内实验。我们发现RPE65蛋白的一个新的45 kDa的截短片段，被命名为RPE45，在光照或氧化应激下出现在RPE细胞中。RPE45是由重组半胱天冬酶通过泛素化依赖机制在体外产生的。总的来说，我们的研究结果表明，视觉周期中的氧化应激导致RPE65的裂解。
The regeneration of the 11-cis-retinyl imine chromophore of rhodopsin during the visual cycle and mechanisms that control this process are central questions in the field of vision research. The retinal pigment epithelium (RPE)-specific protein RPE65 is centrally involved in the isomerization and hydrolysis of alltrans-retinyl esters. In this study, we investigated RPE65 cleavage and potential regulatory mechanisms under oxidative stress conditions. The D407 RPE cell cultures were exposed to H2O2 (100–1000 µM). Changes in the levels of RPE65 and proteins related to apoptosis were investigated using gel electrophoresis and western blotting. Mass spectrometry was used to confirm the identity of RPE65. C57BL/6J (M450) and C3HeB/FeJ (L450) mice were used for in vivo experiments. We found that a novel 45 kDa truncated fragment of the RPE65 protein, designated RPE45, appears in RPE cells upon light exposure or oxidative stress. RPE45 is generated in vitro by recombinant caspases via an ubiquitination-dependent mechanism. Collectively, our results indicate that oxidative stress during the visual cycle results in cleavage of RPE65.
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DOI：
10.1073/pnas.84.7.1849
发表时间：
1987-04-01
影响因子：
11.1
作者：
BERNSTEIN, PS;LAW, WC;RANDO, RR
通讯作者：
RANDO, RR
影响因子：
5.3
作者：
Wenzel, A;Remé, CE;Grimm, C
通讯作者：
Grimm, C
DOI：
10.1073/pnas.0503460102
发表时间：
2005-08-30
影响因子：
11.1
作者：
Moiseyev, G;Chen, Y;Ma, JX
通讯作者：
Ma, JX
影响因子：
6.7
作者：
Hamirally S;Kamil JP;Ndassa-Colday YM;Lin AJ;Jahng WJ;Baek MC;Noton S;Silva LA;Simpson-Holley M;Knipe DM;Golan DE;Marto JA;Coen DM
通讯作者：
Coen DM
影响因子：
2.9
作者：
Jahng, WJ;David, C;Rando, RR
通讯作者：
Rando, RR