title: Tailored Synthesis of 162-Residue S-Monoglycosylated GM2-Activator Protein (GM2AP) Analogues that Allows Facile Access to a Protein Library.
abstract: A synthetic protocol for the preparation of 162-residue S-monoglycosylated GM2-activator protein (GM2AP) analogues bearing various amino acid substitutions for Thr69 has been developed. The facile incorporation of the replacements into the protein was achieved by means of a one-pot/N-to-C-directed sequential ligation strategy using readily accessible middle N-sulfanylethylanilide (SEAlide) peptides each consisting of seven amino acid residues. A kinetically controlled ligation protocol was successfully applied to the assembly of three peptide segments covering the GM2AP. The native chemical ligation (NCL) reactivities of the SEAlide peptides can be tuned by the presence or absence of phosphate salts. Furthermore, NCL of the alkyl thioester fragment [GM2AP (1-31)] with the N-terminal cysteinyl prolyl thioester [GM2AP (32-67)] proceeded smoothly to yield the 67-residue prolyl thioester, with the prolyl thioester moiety remaining intact. This newly developed strategy enabled the facile synthesis of GM2AP analogues. Thus, we refer to this synthetic protocol as "tailored synthesis" for the construction of a GM2AP library.
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Nakamura
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Sato
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Naoto
authors.initials: N
authors.lastname: Naruse
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Kitakaze
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Tsubasa
authors.initials: T
authors.lastname: Inokuma
authors.affiliation: Molecular Profiling Research Center for Drug Discovery, AIST, 2-4-7 Aomi, Koto-ku, Tokyo, 135-0064, Japan.
authors.forename: Takatsugu
authors.initials: T
authors.lastname: Hirokawa
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Akira
authors.initials: A
authors.lastname: Shigenaga
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima, 770-8505, Japan.
authors.forename: Kohji
authors.initials: K
authors.lastname: Itoh
authors.affiliation: Institute of Biomedical Sciences, Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi, Tokushima, 770-8505, Japan. aotaka@tokushima-u.ac.jp.
authors.forename: Akira
authors.initials: A
authors.lastname: Otaka
mesh_terms: D049289:G(M2) Activator Protein / Q000138:chemical synthesis* / Q000737:chemistry; D015394:Molecular Structure; D019151:Peptide Library
chemical_list: D049289:G(M2) Activator Protein; D019151:Peptide Library
keywords: GM2AP; chemoselectivity; kinetically controlled ligation; native chemical ligation; protein modifications