Aromatic thiol-mediated cleavage of N-O bonds enables chemical ubiquitylation of folded proteins.
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.
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Caroline E
CE
Weller
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Abhinav
A
Dhall
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Feizhi
F
Ding
Departamento de Bioquímica, Instituto de Química-Universidade de São Paulo, São Paulo 05513-970, Brazil.
Edlaine
E
Linares
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Samuel D
SD
Whedon
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Nicholas A
NA
Senger
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Elizabeth L
EL
Tyson
Department of Chemistry, Princeton University, Princeton, New Jersey 08544, USA.
John D
JD
Bagert
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Xiaosong
X
Li
Departamento de Bioquímica, Instituto de Química-Universidade de São Paulo, São Paulo 05513-970, Brazil.
Ohara
http://orcid.org/0000-0002-7220-4286
O
Augusto
Department of Chemistry, University of Washington, Seattle, Washington 98195, USA.
Champak
C
Chatterjee
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