Construction of highly glycosylated mucin-type glycopeptides based on microwave-assisted solid-phase syntheses and enzymatic modifications.
A MUC1-related glycopeptide having five core-2 hexasaccharide branches (C330H527N46O207, MW = 8450.9) was synthesized by a new strategy using a combination of microwave-assisted solid-phase synthesis (MA-SPGS) and enzymatic sugar elongation. Synthesis of a key glycopeptide intermediate was best achieved in a combination of PEGA [poly(ethylene glycol)-poly-(N,N-dimethylacrylamide) copolymer] resin and MA-SPGS using glycosylated amino acid building blocks with high speed and high purity. Deprotection of the glycopeptide intermediate and subsequent glycosyltransferase-catalyzed sugar elongations were performed for generation of the additional diversities with the sugar moieties of glycopeptides using beta1,4-galactosyltransferase (beta1,4-GalT) and two kinds of alpha2,3-sialyltransferases [ST3Gal III; alpha2,3-(N)-SiaT and ST3Gal II; alpha2,3-(O)-SiaT]. These reactions proceeded successfully in the presence of 0.2% Triton X-100 to convert the chemically synthesized trisaccharide glycans to disialylated hexasaccharide.
Matsushita
Takahiko
T
Division of Biological Sciences, Graduate School of Science, Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, Sapporo, 001-0021 Japan.
Hinou
Hiroshi
H
Fumoto
Masataka
M
Kurogochi
Masaki
M
Fujitani
Naoki
N
Shimizu
Hiroki
H
Nishimura
Shin-ichiro
S
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry; D002236:Carbohydrate Conformation; D002851:Chromatography, High Pressure Liquid; D006020:Glycopeptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D006031:Glycosylation; D016695:Glycosyltransferases / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D008872:Microwaves; D008969:Molecular Sequence Data; D009077:Mucins / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000737:chemistry; D011137:Polystyrenes / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000596:Amino Acids; D006020:Glycopeptides; D009077:Mucins; D011137:Polystyrenes; C101297:Tentagel resin; C109511:bis(polyoxyethylene bis(amine)); D011092:Polyethylene Glycols; D016695:Glycosyltransferases