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PreprintCited by 21Redox biology and oxidative stressAdvanced Proteomics Techniques and ApplicationsClick Chemistry and Applications

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Defining the Cell Surface Cysteinome using Two-step Enrichment Proteomics

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Yan, Tianyang, Boatner, Lisa M., Cui, Liujuan, Tontonoz, Peter, Backus, Keriann M.·2023·JACS Au
DOI 10.1101/2023.10.17.562832PMC PMC10751780

Preprint — published as 10.1021/jacsau.3c00707

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Abstract

The plasma membrane proteome is a rich resource of functional 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 Surf ace Cys teinome. 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 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.

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Metadata

Authors
Yan, T., Boatner, L. M., Cui, L., Tontonoz, P., & Backus, K. M.
Affiliations
University of California, Los Angeles (US)
Corresponding author
Backus, Keriann M.
ORCID
Venue
JACS Au
Publisher
openRxiv
Published
2023-10-19
Volume
3
Issue
12
Pages
3506-3523
ISSN
2691-3704 (ISSN-L)
DOI
10.1101/2023.10.17.562832
PMC
PMC10751780
OpenAlex
W4389663659
Cited by
21
References
1
Citations by year
2026: 3 · 2025: 8 · 2024: 9 · 2023: 1
FWCI
1.87
Type
journal article
Language
en
Open access
Licence
cc-by-nc-nd
Locations
  • JACS Au (publishedVersion, cc-by-nc-nd)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • eScholarship (California Digital Library) (submittedVersion, cc-by-nc-nd)
  • PubMed Central (submittedVersion, cc-by-nc-nd)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • David and Lucile Packard Foundation
  • Tobacco-Related Disease Research Program (T31DT1800)
  • Division of Intramural Research, National Institute of Allergy and Infectious Diseases (5P30 AI028697)
  • National Institutes of Health (DP2 OD030950-01, AI028697)
  • National Heart, Lung, and Blood Institute (P01HL146358)
  • NIH Office of the Director (DP2 OD030950-01)
  • National Institute of General Medical Sciences (T32GM008185)
  • National Institute of Allergy and Infectious Diseases (P30AI028697, P30AI152501)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
Keywords
plasma membrane proteome, LDL receptor, LDL uptake, protein localization

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