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A clickable probe for versatile characterization of S-nitrosothiols.

ABSTRACT: S-nitrosation of cysteine thiols (SNOs), commonly referred to as S-nitrosylation, is a cysteine oxoform that plays an important role in cellular signaling and impacts protein function and stability. Direct labeling of SNOs in cells with the flexibility to perform a wide range of cellular and biochemical assays remains a bottleneck as all SNO-targeted probes to date employ a single analytical modality such as biotin or a specific fluorophore. We therefore developed a clickable, alkyne-containing SNO probe 'PBZyn' based on the o-phosphino-benzoyl group warhead that enables multi-modal analysis via click conjugation. We demonstrate the utility of PBZyn to assay SNOs using in situ cellular imaging, protein blotting and affinity purification, as well as mass spectrometry. The flexible PBZyn probe will greatly facilitate investigation into the regulation of SNOs.

SUBMITTER: Clements JL

PROVIDER: S-EPMC7490559 | biostudies-literature | 2020 Sep

REPOSITORIES: biostudies-literature

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A clickable probe for versatile characterization of S-nitrosothiols.

Clements Jenna L JL Pohl Franziska F Muthupandi Pandi P Rogers Stephen C SC Mao Jack J Doctor Allan A Birman Vladimir B VB Held Jason M JM

Redox biology 20200901

S-nitrosation of cysteine thiols (SNOs), commonly referred to as S-nitrosylation, is a cysteine oxoform that plays an important role in cellular signaling and impacts protein function and stability. Direct labeling of SNOs in cells with the flexibility to perform a wide range of cellular and biochemical assays remains a bottleneck as all SNO-targeted probes to date employ a single analytical modality such as biotin or a specific fluorophore. We therefore developed a clickable, alkyne-containing ...[more]

PMID: 32916549

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