title: One-pot chemical synthesis of small ubiquitin-like modifier protein-peptide conjugates using bis(2-sulfanylethyl)amido peptide latent thioester surrogates.
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.
authors.affiliation: Centre National de la Recherche Scientifique, Lille Institute of Biology, Institut Pasteur de Lille, University of Lille, Lille, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Boll
authors.affiliation: Centre National de la Recherche Scientifique, Lille Institute of Biology, Institut Pasteur de Lille, University of Lille, Lille, France.
authors.forename: Hervé
authors.initials: H
authors.lastname: Drobecq
authors.affiliation: Centre National de la Recherche Scientifique, Lille Institute of Biology, Institut Pasteur de Lille, University of Lille, Lille, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Ollivier
authors.affiliation: Centre National de la Recherche Scientifique, Lille Institute of Biology, Institut Pasteur de Lille, University of Lille, Lille, France.
authors.forename: Annick
authors.initials: A
authors.lastname: Blanpain
authors.affiliation: Centre National de la Recherche Scientifique, Lille Institute of Biology, Institut Pasteur de Lille, University of Lille, Lille, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Raibaut
authors.affiliation: Centre National de la Recherche Scientifique, Lille Institute of Biology, Institut Pasteur de Lille, University of Lille, Lille, France.
authors.forename: Rémi
authors.initials: R
authors.lastname: Desmet
authors.affiliation: Centre National de la Recherche Scientifique, Lille Institute of Biology, Institut Pasteur de Lille, University of Lille, Lille, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Vicogne
authors.affiliation: Centre National de la Recherche Scientifique, Lille Institute of Biology, Institut Pasteur de Lille, University of Lille, Lille, France.
authors.forename: Oleg
authors.initials: O
authors.lastname: Melnyk
mesh_terms: D000595:Amino Acid Sequence; D060326:Chemistry Techniques, Synthetic; D005449:Fluorenes; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D060327:Solid-Phase Synthesis Techniques; D058207:Sumoylation; D025801:Ubiquitin / Q000737:chemistry*
chemical_list: C496789:9-fluorenylmethoxycarbonyl; D005449:Fluorenes; D010455:Peptides; D025801:Ubiquitin