From the Laboratory of Translational Neuropharmacology and.
J Biol Chem. 2014 Feb 21;289(8):4660-73. doi: 10.1074/jbc.M113.510883. Epub 2013 Dec 26.
The subcellular distribution of the G protein-coupled receptor GPR37 affects cell viability and is implicated in the pathogenesis of parkinsonism. Intracellular accumulation and aggregation of GPR37 cause cell death, whereas GPR37 located in the plasma membrane provides cell protection. We define here a pathway through which the recently identified natural ligand, prosaposin, promotes plasma membrane association of GPR37. Immunoabsorption of extracellular prosaposin reduced GPR37(tGFP) surface density and decreased cell viability in catecholaminergic N2a cells. We found that GPR37(tGFP) partitioned in GM1 ganglioside-containing lipid rafts in the plasma membrane of live cells. This partitioning required extracellular prosaposin and was disrupted by lipid raft perturbation using methyl-β-cyclodextrin or cholesterol oxidase. Moreover, complex formation between GPR37(tGFP) and the GM1 marker cholera toxin was observed in the plasma membrane. These data show functional association between GPR37, prosaposin, and GM1 in the plasma membrane. These results thus tie together the three previously defined components of the cellular response to insult. Our findings identify a mechanism through which the receptor's natural ligand and GM1 may protect against toxic intracellular GPR37 aggregates observed in parkinsonism.
G 蛋白偶联受体 GPR37 的亚细胞分布影响细胞活力,并与帕金森病的发病机制有关。GPR37 的细胞内积累和聚集导致细胞死亡,而位于质膜中的 GPR37 则提供细胞保护。我们在这里定义了一条途径,即最近发现的天然配体脑苷脂激活蛋白通过该途径促进 GPR37 与质膜的结合。细胞外脑苷脂激活蛋白的免疫吸收降低了儿茶酚胺能 N2a 细胞中 GPR37(tGFP)的表面密度,并降低了细胞活力。我们发现 GPR37(tGFP)在活细胞质膜中的 GM1 神经节苷脂包含的脂筏中进行分区。这种分区需要细胞外脑苷脂激活蛋白,并且可以通过使用甲基-β-环糊精或胆固醇氧化酶破坏脂筏来破坏。此外,在质膜中观察到 GPR37(tGFP)与 GM1 标记霍乱毒素之间的复合物形成。这些数据显示 GPR37、脑苷脂激活蛋白和 GM1 在质膜中具有功能关联。这些结果将帕金森病中观察到的细胞对损伤的三种先前定义的反应成分联系在一起。我们的发现确定了一种机制,通过该机制,受体的天然配体和 GM1 可能防止帕金森病中观察到的有毒细胞内 GPR37 聚集。