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Profiling Protein S-Sulfination with Maleimide-Linked Probes.

ABSTRACT: Cysteine residues are susceptible to oxidation to form S-sulfinyl (R-SO2 H) and S-sulfonyl (R-SO3 H) post-translational modifications. Here we present a simple bioconjugation strategy to label S-sulfinated proteins by using reporter-linked maleimides. After alkylation of free thiols with iodoacetamide, S-sulfinated cysteines react with maleimide to form a sulfone Michael adduct that remains stable under acidic conditions. Using this sequential alkylation strategy, we demonstrate differential S-sulfination across mouse tissue homogenates, as well as enhanced S-sulfination following pharmacological induction of endoplasmic reticulum stress, lipopolysaccharide stimulation, and inhibitors of the electron transport chain. Overall, this study reveals a broadened profile of maleimide reactivity across cysteine modifications, and outlines a simple method for profiling the physiological role of cysteine S-sulfination in disease.

SUBMITTER: Kuo YH

PROVIDER: S-EPMC5765543 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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Profiling Protein S-Sulfination with Maleimide-Linked Probes.

Kuo Yu-Hsuan YH Konopko Aaron M AM Borotto Nicholas B NB Majmudar Jaimeen D JD Haynes Sarah E SE Martin Brent R BR

Chembiochem : a European journal of chemical biology 20170901 20

Cysteine residues are susceptible to oxidation to form S-sulfinyl (R-SO2 H) and S-sulfonyl (R-SO3 H) post-translational modifications. Here we present a simple bioconjugation strategy to label S-sulfinated proteins by using reporter-linked maleimides. After alkylation of free thiols with iodoacetamide, S-sulfinated cysteines react with maleimide to form a sulfone Michael adduct that remains stable under acidic conditions. Using this sequential alkylation strategy, we demons ...[more]

PMID: 28809078

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