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Integrating cell surface cysteine enrichment chemoproteomics, subcellular localization annotations and experimental oxidation measurements to define the cell surface cysteinome.

License: Apache License 2.0

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CysSurfDB: Cell Surface Cysteinome Localization Database

Integrating cell surface cysteine enrichment chemoproteomics, subcellular localization annotations and experimental oxidation measurements to define the cell surface cysteinome. Available at: https://pubs.acs.org/doi/10.1021/jacsau.3c00707

The CysSurf Database and mass spectrometry processing were created and developed by Lisa Boatner.

About: The plasma membrane proteome is a rich resource of functionally important and therapeutically relevant protein targets. Distinguished by high hydrophobicity, heavy glycosylation, disulfide-rich sequences, and low overall abundance, the cell surface proteome remains undersampled in established proteomic pipelines, including our own cysteine chemoproteomics platforms. Here, we paired cell surface glycoprotein capture with cysteine chemoproteomics to establish a two-stage enrichment method that enables chemoproteomic profiling of cell Surface Cysteinome. Our “Cys-Surf” platform captures >2,800 total membrane protein cysteines in 1,046 proteins, including 1,907 residues not previously captured by bulk proteomic analysis. By pairing Cys-Surf with an isotopic chemoproteomic readout, we uncovered 821 total ligandable cysteines, including known and novel sites. Cys-Surf also robustly delineates redox-sensitive cysteines, including cysteines prone to activation-dependent changes to cysteine oxidation state and residues sensitive to addition of exogenous reductants. Exemplifying the capacity of Cys-Surf to delineate functionally important cysteines, we identified a redox sensitive cysteine in the low-density lipoprotein receptor (LDLR) that impacts both the protein localization and uptake of low-density lipoprotein (LDL) particles. Taken together, the Cys-Surf platform, distinguished by its two-stage enrichment paradigm, represents a tailored approach to delineate the functional and therapeutic potential of the plasma membrane cysteinome.

Yan, T., Boatner, L. M., Cui, L., Tontonoz, P., & Backus, K. M. (2023). Defining the Cell Surface Cysteinome using Two-step Enrichment Proteomics. JACS Au.

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