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Vesicle-associated membrane protein 2 is a cargo-selective v-SNARE for a subset of GPCRs
The Journal of Cell Biology Pub Date : 2023-04-06 , DOI: 10.1083/jcb.202207070
Hao Chen 1 , Zara Y Weinberg 1 , G Aditya Kumar 1 , Manojkumar A Puthenveedu 1
Affiliation  

Vesicle fusion at the plasma membrane is critical for releasing hormones and neurotransmitters and for delivering the cognate G protein–coupled receptors (GPCRs) to the cell surface. The SNARE fusion machinery that releases neurotransmitters has been well characterized. In contrast, the fusion machinery that delivers GPCRs is still unknown. Here, using high-speed multichannel imaging to simultaneously visualize receptors and v-SNAREs in real time in individual fusion events, we identify VAMP2 as a selective v-SNARE for GPCR delivery. VAMP2 was preferentially enriched in vesicles that mediate the surface delivery of μ opioid receptor (MOR), but not other cargos, and was required selectively for MOR recycling. Interestingly, VAMP2 did not show preferential localization on MOR-containing endosomes, suggesting that v-SNAREs are copackaged with specific cargo into separate vesicles from the same endosomes. Together, our results identify VAMP2 as a cargo-selective v-SNARE and suggest that surface delivery of specific GPCRs is mediated by distinct fusion events driven by distinct SNARE complexes.

中文翻译:

囊泡相关膜蛋白 2 是 GPCR 子集的货物选择性 v-SNARE

质膜上的囊泡融合对于释放激素和神经递质以及将同源 G 蛋白偶联受体 (GPCR) 传递到细胞表面至关重要。释放神经递质的 SNARE 融合机制已经得到了很好的表征。相比之下,传递 GPCR 的融合机制仍然未知。在这里,我们使用高速多通道成像同时实时可视化单个融合事件中的受体和 v-SNARE,我们将 VAMP2 确定为用于 GPCR 传递的选择性 v-SNARE。 VAMP2 优先富集在介导 μ 阿片受体 (MOR) 表面递送的囊泡中,但不富集在其他货物中,并且是 MOR 回收所选择性需要的。有趣的是,VAMP2 没有显示出在包含 MOR 的内体上的优先定位,这表明 v-SNARE 与特定货物一起包装到来自相同内体的单独囊泡中。总之,我们的结果将 VAMP2 确定为货物选择性 v-SNARE,并表明特定 GPCR 的表面递送是由不同 SNARE 复合物驱动的不同融合事件介导的。
更新日期:2023-04-06
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