Chemoenzymatic synthesis of neoglycoproteins using transglycosylation with endo-beta-N-acetylglucosaminidase A.
A novel chemoenzymatic approach to synthesize neoglycoproteins containing high-mannose-type oligosaccharides is described. p-Isothiocyanatophenyl-beta-d-glucopyranoside (Glc-ITC) was transferred to the reducing end of the high-mannose-type oligosaccharides using a transglycosylation activity of endo-beta-N-acetylglucosaminidase A (Endo-A). A novel oligosaccharide, Man(6)GlcNAc-Glc-ITC, was synthesized as a coupling reagent for lysyl and N-terminal residues of the protein moiety. The neoglycoconjugate was coupled with several nonglycosylated proteins such as ribonuclease A, lysozyme, and alpha-lactalbumin. Between one and four high-mannose-type oligosaccharides were incorporated per molecule of these proteins. This method should be very useful for the synthesis of neoglycoproteins with homogeneous high-mannose-type oligosaccharides.
Fujita
K
K
Department of Life Sciences, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan.
Takegawa
K
K
D002240:Carbohydrate Sequence; D006001:Glycoconjugates / Q000096:biosynthesis / Q000138:chemical synthesis / Q000737:chemistry; D006023:Glycoproteins / Q000096:biosynthesis* / Q000138:chemical synthesis* / Q000737:chemistry; D006031:Glycosylation; D066298:In Vitro Techniques; D008358:Mannose / Q000737:chemistry; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / Q000378:metabolism*; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000096:biosynthesis / Q000138:chemical synthesis / Q000737:chemistry
D006001:Glycoconjugates; D006023:Glycoproteins; D009844:Oligosaccharides; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D008358:Mannose