Premium
Molecular Design of Glycoprotein Mimetics: Glycoblotting by Engineered Proteins with an Oxylamino‐Functionalized Amino Acid Residue
Author(s) -
Matsubara Naoki,
Oiwa Kei,
Hohsaka Takahiro,
Sadamoto Reiko,
Niikura Kenichi,
Fukuhara Norio,
Takimoto Akio,
Kondo Hirosato,
Nishimura ShinIchiro
Publication year - 2005
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.200500531
Subject(s) - glycosylation , glycoprotein , biochemistry , chemistry , streptavidin , protein engineering , translation (biology) , amino acid , residue (chemistry) , escherichia coli , biotin , gene , messenger rna , enzyme
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.