Structure and function in rhodopsin: Mass spectrometric identification of the abnormal intradiscal disulfide bond in misfolded retinitis pigmentosa mutants
Structure and function in rhodopsin: Mass spectrometric identification of the abnormal intradiscal disulfide bond in misfolded retinitis pigmentosa mutants
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DOI：
10.1073/pnas.061632798
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发表时间：
2001-04-24
影响因子：
11.1
通讯作者：
Khorana, HG
中科院分区：
文献类型：
--
作者：
Hwa, J;Klein-Seetharaman, J;Khorana, HG
视网膜色素性视网膜炎（RP）点突变在视网膜内（ID）和跨膜区域的视紫红质导致部分或完全错误折叠视紫红质。导致il -顺式视网膜结合丧失。先前的研究表明，错误折叠是由于在天然视紫质中形成的与天然Cys-110-Cys-187二硫键不同的ID结构域的二硫键引起的。在这里，我们报告了通过质谱分析直接鉴定跨膜区域错误折叠的RP突变体的异常二硫键。这个二硫键在Cys-185和Cys-187之间。与之前在ID结构域错误折叠的RP突变中发现的相同。这里描述的策略应该普遍适用于鉴定其他完整膜蛋白中的二硫键。
Retinitis pigmentosa (RP) point mutations in both the intradiscal (ID) and transmembrane domains of rhodopsin cause partial or complete misfolding of rhodopsin. resulting in loss of Il-cis-retinal binding. Previous work has shown that misfolding is caused by the formation of a disulfide bond in the ID domain different from the native Cys-110-Cys-187 disulfide bond in native rhodopsin. Here we report on direct identification of the abnormal disulfide bond in misfolded RP mutants in the transmembrane domain by mass spectrometric analysis. This disulfide bond is between Cys-185 and Cys-187. the same as previously identified in misfolded RP mutations in the ID domain. The strategy described here should be generally applicable to identification of disulfide bonds in other integral membrane proteins.