Multiplexed proteomic profiling of cysteine reactivity and ligandability in human T cells

Ekaterina V Vinogradova; Benjamin F Cravatt

doi:10.1016/j.xpro.2021.100458

Multiplexed proteomic profiling of cysteine reactivity and ligandability in human T cells

STAR Protoc. 2021 Apr 8;2(2):100458. doi: 10.1016/j.xpro.2021.100458. eCollection 2021 Jun 18.

Authors

Ekaterina V Vinogradova^{1

2}, Benjamin F Cravatt¹

Affiliations

¹ Department of Chemistry, The Scripps Research Institute, La Jolla, CA 92037, USA.
² Laboratory of Chemical Immunology and Proteomics, The Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.

Abstract

Differential amino acid reactivity with chemical probes can provide valuable information on the functionality and ligandability of proteins in native biological systems. Here, we present a quantitative, multiplexed chemical proteomic protocol for in-depth reactivity and ligandability profiling of cysteines in proteins in quiescent and stimulated T cells. This protocol illuminates dynamic immune state-dependent alterations in cysteine reactivity, revealing chemoselective and stereoselective small-molecule interactions with cysteines in structurally and functionally diverse proteins that lack chemical probes. For complete details on the use and execution of this protocol, please refer to Vinogradova et al. (2020).

Keywords: Cell isolation; Flow Cytometry/Mass Cytometry; High Throughput Screening; Immunology; Molecular/Chemical Probes; Protein Biochemistry; Proteomics.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cysteine / metabolism*
Humans
Proteome / metabolism*
Proteomics*
T-Lymphocytes / cytology
T-Lymphocytes / metabolism*

Substances

Proteome
Cysteine

Grants and funding

R35 CA231991/CA/NCI NIH HHS/United States