Molecular design of glycoprotein mimetics: glycoblotting by engineered proteins with an oxylamino-functionalized amino acid residue.
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.
Matsubara
Naoki
N
Laboratory for Bio-Macromolecular Chemistry, Division of Biological Sciences, Hokkaido University, Sapporo 001-0021, Japan.
Oiwa
Kei
K
Hohsaka
Takahiro
T
Sadamoto
Reiko
R
Niikura
Kenichi
K
Fukuhara
Norio
N
Takimoto
Akio
A
Kondo
Hirosato
H
Nishimura
Shin-Ichiro
S
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
D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins