Details

Autor(en) / Beteiligte

• Weller, Caroline E.
• Dhall, Abhinav
• Ding, Feizhi
• Linares, Edlaine
• Whedon, Samuel D.
• Senger, Nicholas A.
• Tyson, Elizabeth L.
• Bagert, John D.
• Li, Xiaosong
• Augusto, Ohara
• Chatterjee, Champak

Titel

Aromatic thiol-mediated cleavage of N–O bonds enables chemical ubiquitylation of folded proteins

Ist Teil von

• Nature communications, 2016-09, Vol.7 (1), p.12979-12979, Article 12979

Ort / Verlag

London: Nature Publishing Group UK

Erscheinungsjahr

2016

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• Access to protein substrates homogenously modified by ubiquitin (Ub) is critical for biophysical and biochemical investigations aimed at deconvoluting the myriad biological roles for Ub. Current chemical strategies for protein ubiquitylation, however, employ temporary ligation auxiliaries that are removed under harsh denaturing conditions and have limited applicability. We report an unprecedented aromatic thiol-mediated N–O bond cleavage and its application towards native chemical ubiquitylation with the ligation auxiliary 2-aminooxyethanethiol. Our interrogation of the reaction mechanism suggests a disulfide radical anion as the active species capable of cleaving the N–O bond. The successful semisynthesis of full-length histone H2B modified by the small ubiquitin-like modifier-3 (SUMO-3) protein further demonstrates the generalizability and compatibility of our strategy with folded proteins. Chemical approaches to site-specifically ubiquitylate a target protein allow investigation of the biochemical effects of this modification, but they often destabilize the protein. Here, the authors report on a synthetic conjugation strategy that leads to protein ubiquitylation in non-denaturing conditions.