Abstract:
:Small ubiquitin-like modifier (SUMO) post-translational modification (PTM) of proteins has a crucial role in the regulation of important cellular processes. This protocol describes the chemical synthesis of functional SUMO-peptide conjugates. The two crucial stages of this protocol are the solid-phase synthesis of peptide segments derivatized by thioester or bis(2-sulfanylethyl)amido (SEA) latent thioester functionalities and the one-pot assembly of the SUMO-peptide conjugate by a sequential native chemical ligation (NCL)/SEA native peptide ligation reaction sequence. This protocol also enables the isolation of a SUMO SEA latent thioester, which can be attached to a target peptide or protein in a subsequent step. It is compatible with 9-fluorenylmethoxycarbonyl (Fmoc) chemistry, and it gives access to homogeneous, reversible and functional SUMO conjugates that are not easily produced using living systems. The synthesis of SUMO-peptide conjugates on a milligram scale takes 20 working days.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Boll E,Drobecq H,Ollivier N,Blanpain A,Raibaut L,Desmet R,Vicogne J,Melnyk Odoi
10.1038/nprot.2015.013subject
Has Abstractpub_date
2015-02-01 00:00:00pages
269-92issue
2eissn
1754-2189issn
1750-2799pii
nprot.2015.013journal_volume
10pub_type
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