A clickable probe for versatile characterization of S-nitrosothiols
AuthorClements, Jenna L.
Rogers, Stephen C.
Birman, Vladimir B.
Held, Jason M.
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AbstractS-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.
SponsorsWashington University Research Strategic Alliance
Rights/Terms© 2020 Published by Elsevier B.V.
Identifier to cite or link to this itemhttp://hdl.handle.net/10713/13709