TY - JOUR
T1 - Photoproximity Labeling of Sialylated Glycoproteins (GlycoMap) Reveals Sialylation-Dependent Regulation of Ion Transport
AU - Meyer, Claudio F.
AU - Seath, Ciaran P.
AU - Knutson, Steve D.
AU - Lu, Wenyun
AU - Rabinowitz, Joshua D.
AU - Macmillan, David W.C.
N1 - Funding Information:
This work was funded by the NIH National Institute of General Medical Sciences (R35-GM134897-02) and kind gifts from Merck, BMS, Pfizer, Janssen, Genentech, and Eli Lilly. The authors also acknowledge the Princeton Catalysis Initiative and the Ludwig Cancer Research for supporting this work. They also acknowledge the Swiss National Science Foundation (P2SKP2_199458, C.F.M.) and the NIH (1F32GM142206-01, S.D.K.; R50CA211437, W.L.) for fellowships. The authors thank Saw Kyin and Henry H. Shwe at the Princeton Proteomics Facility. The authors acknowledge the use of Princeton’s Imaging and Analysis Center, which is partially supported by the Princeton Center for Complex Materials, National Science Foundation/Materials Research Science and Engineering Centers program (DMR-1420541). Generalized schemes were created using Biorender.
Publisher Copyright:
© 2022 American Chemical Society.
PY - 2022/12/28
Y1 - 2022/12/28
N2 - 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.
AB - 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.
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U2 - 10.1021/jacs.2c11094
DO - 10.1021/jacs.2c11094
M3 - Article
C2 - 36525649
AN - SCOPUS:85144350372
SN - 0002-7863
VL - 144
SP - 23633
EP - 23641
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 51
ER -