R17C Mutation in Photoreceptor Disc-Specific Protein, PRCD, Results in Additional Lipidation Altering Protein Stability and Subcellular Localization.
R17C Mutation in Photoreceptor Disc-Specific Protein, PRCD, Results in Additional Lipidation Altering Protein Stability and Subcellular Localization.
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DOI：
10.3390/ijms231810802
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发表时间：
2022-09-16
影响因子：
5.6
通讯作者：
Kolandaivelu, Saravanan
中科院分区：
文献类型：
--
作者：
Myers, Boyden;Sechrest, Emily R.;Hamner, Gabrielle;Motipally, Sree, I;Murphy, Joseph;Kolandaivelu, Saravanan
关键词：
进行性视杆-视锥变性（PRCD）是光感受器外节（OS）盘特异性蛋白，其对于维持OS结构必不可少，同时有助于视紫红质包装密度和在盘膜中的分布。以前，我们显示PRCD在唯一的半胱氨酸（Cys 2），其中与视网膜色素变性（RP）在人类和狗的突变链接棕榈酰化证明了蛋白质的稳定性和贩运到OS的重要性。我们证明了一个突变，在PRCD（Arg 17 Cys）与RP连接的多碱区（PBR），通过酰基RAC观察到额外的脂化。hRPE 1细胞中瞬时表达的R17 C的免疫定位描绘了与野生型PRCD相似的特征;然而，在Cys 2 Tyr处缺乏内源性棕榈酰化的双突变体与Arg 17 Cys与C2 Y蛋白相当，因为两者都聚集，错误定位于细胞质内的亚细胞区室。视网膜下注射PRCD突变体构建体，然后在鼠视网膜中电穿孔，表现出在内段中的错误定位。尽管被另外脂化并表现出强的膜缔合，但PBR中的突变影响蛋白质稳定性和定位于OS。单独的PBR内的酰化既不补偿蛋白质稳定性也不补偿运输，揭示了PBR中的缺陷可能导致与致盲疾病相关的PRCD蛋白的失调。
Progressive rod-cone degeneration (PRCD) is a photoreceptor outer segment (OS) disc-specific protein essential for maintaining OS structures while contributing to rhodopsin packaging densities and distribution in disc membranes. Previously, we showed PRCD undergoing palmitoylation at the sole cysteine (Cys2), where a mutation linked with retinitis pigmentosa (RP) in humans and dogs demonstrates the importance of palmitoylation for protein stability and trafficking to the OS. We demonstrate a mutation, in the polybasic region (PBR) of PRCD (Arg17Cys) linked with RP where an additional lipidation is observed through acyl-RAC. Immunolocalization of transiently expressed R17C in hRPE1 cells depicts similar characteristics to wild-type PRCD; however, a double mutant lacking endogenous palmitoylation at Cys2Tyr with Arg17Cys is comparable to the C2Y protein as both aggregate, mislocalized to the subcellular compartments within the cytoplasm. Subretinal injection of PRCD mutant constructs followed by electroporation in murine retina exhibit mislocalization in the inner segment. Despite being additionally lipidated and demonstrating strong membrane association, the mutation in the PBR affects protein stability and localization to the OS. Acylation within the PBR alone neither compensates for protein stability nor trafficking, revealing defects in the PBR likely lead to dysregulation of PRCD protein associated with blinding diseases.
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