Photoproximity Labeling of Sialylated Glycoproteins (GlycoMap) Reveals Sialylation-Dependent Regulation of Ion Transport.
Photoproximity Labeling of Sialylated Glycoproteins (GlycoMap) Reveals Sialylation-Dependent Regulation of Ion Transport.
复制标题
唾液酸化糖蛋白(GlycoMap)的光邻近标记揭示了依赖唾液酸化的离子转运调节。
DOI：
10.1021/jacs.2c11094
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发表时间：
2022-12
影响因子：
15
通讯作者：
Claudio F. Meyer;Ciaran P. Seath;Steve D. Knutson;Wenyun Lu;J. Rabinowitz;D. MacMillan
中科院分区：
文献类型：
--
作者：
Claudio F. Meyer;Ciaran P. Seath;Steve D. Knutson;Wenyun Lu;J. Rabinowitz;D. MacMillan
唾液酸化是一种关键的蛋白质翻译后修饰，有助于神经发育、肿瘤发生和免疫反应。在癌症中，唾液酸化显著上调。然而，唾液酸化的功能生化后果仍然是个谜。在这里，我们建立了一个μ图谱邻近标记平台，它利用代谢性插入的叠氮唾液酸光催化剂在唾液酸化的细胞表面糖蛋白上引入Ir光催化剂，作为描述唾液酸化蛋白质组局部微环境的一种手段。在原代宫颈细胞和癌细胞系(HeLa)之间的比较实验中，我们确定了全局唾液体和近端蛋白质的关键差异，包括调节代谢物和离子运输的溶质载体蛋白质。特别是，我们发现转运乙醇胺和锌的受体之间的细胞表面相互作用是唾液酸化依赖的，并影响细胞内代谢物的水平。这些结果建立了一种询问蛋白多糖功能的μ图谱方法，并支持唾液酸糖蛋白在调节细胞表面转运蛋白中的作用。
Sialylation, the addition of sialic acid to glycans, is a crucial post-translational modification of proteins, contributing to neurodevelopment, oncogenesis, and immune response. In cancer, sialylation is dramatically upregulated. Yet, the functional biochemical consequences of sialylation remain mysterious. Here, we establish a μMap proximity labeling platform that utilizes metabolically inserted azidosialic acid to introduce iridium-based photocatalysts on sialylated cell-surface glycoproteins as a means to profile local microenvironments across the sialylated proteome. In comparative experiments between primary cervical cells and a cancerous cell line (HeLa), we identify key differences in both the global sialome and proximal proteins, including solute carrier proteins that regulate metabolite and ion transport. In particular, we show that cell-surface interactions between receptors trafficking ethanolamine and zinc are sialylation-dependent and impact intracellular metabolite levels. These results establish a μMap method for interrogating proteoglycan function and support a role for sialylated glycoproteins in regulating cell-surface transporters.