title: Molecular design of glycoprotein mimetics: glycoblotting by engineered proteins with an oxylamino-functionalized amino acid residue.
abstract: The general and efficient method for the site-directed glycosylation of proteins is a key step in order to understand the biological importance of the carbohydrate chains of proteins and to control functional roles of the engineered glycoproteins in terms of the development of improved glycoprotein therapeutics. We have developed a novel method for site-directed glycosylation of proteins based on chemoselective blotting of common reducing sugars by genetically encoded proteins. The oxylamino-functionalized L-homoserine residues, 2-amino-4-O-(N-methylaminooxy) butanoic acid and 2-amino-4-aminooxy butanoic acid, were efficiently incorporated into proteins by using the four-base codon/anticodon pair strategy in Escherichia coli in vitro translation. Direct and chemoselective coupling between unmodified simple sugars and N-methylaminooxy group displayed on the engineered streptavidin allowed for the combinatorial synthesis of novel glycoprotein mimetics.
authors.affiliation: Laboratory for Bio-Macromolecular Chemistry, Division of Biological Sciences, Hokkaido University, Sapporo 001-0021, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Matsubara
authors.forename: Kei
authors.initials: K
authors.lastname: Oiwa
authors.forename: Takahiro
authors.initials: T
authors.lastname: Hohsaka
authors.forename: Reiko
authors.initials: R
authors.lastname: Sadamoto
authors.forename: Kenichi
authors.initials: K
authors.lastname: Niikura
authors.forename: Norio
authors.initials: N
authors.lastname: Fukuhara
authors.forename: Akio
authors.initials: A
authors.lastname: Takimoto
authors.forename: Hirosato
authors.initials: H
authors.lastname: Kondo
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry*; D002846:Chromatography, Affinity; D002855:Chromatography, Thin Layer; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry*; D006031:Glycosylation; D013058:Mass Spectrometry; D018716:Molecular Mimicry; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D015202:Protein Engineering; D016339:Spectrometry, Mass, Fast Atom Bombardment
chemical_list: D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins