Enzymatic synthesis of the core-2 sialyl Lewis X O-glycan on the tumor-associated MUC1a' peptide.
Starting from a tumor-associated synthetic MUC1-derived peptide MUC1a' and using a completely enzymatic approach for the synthesis of the core-2 sialyl Lewis X glycopart, the following glycopeptide was synthesized: AHGV[Neu5Ac(alpha2-3)Gal(beta1-4)[Fuc(alpha1-3)]GlcNAc(beta1-6)[Gal(beta1-3)]GalNAc(alpha1-O)]TSAPDTR. First, polypeptide N-acetylgalactosaminyltransferase 3 was used to site-specifically glycosylate MUC1a' to give MUC1a'-GalNAc. Then, in a one-pot reaction employing beta-galactosidase and core-2 beta6-N-acetylglucosaminyltransferase the core-2 O-glycan structure was prepared. The core-2 structure was then sequentially galactosylated, sialylated, and fucosylated by making use of beta4-galactosyltransferase 1, alpha3-sialyltransferase 3, and alpha3-fucosyltransferase 3, respectively, resulting in the sialyl Lewis X glycopeptide. The overall yield of the final compound was 23% (3.2 mg, 1.4 micromol). During the synthesis three intermediate glycopeptides containing O-linked GalNAc, Gal(beta1-4)GlcNAc(beta1-6)[Gal(beta1-3)]GalNAc, and Neu5Ac(alpha2-3)Gal(beta1-4)GlcNAc(beta1-6)[Gal(beta1-3)]GalNAc, respectively, were isolated in mg quantities. All products were characterized by mass spectrometry and NMR spectroscopy.
Gutiérrez Gallego
Ricardo
R
Section of Glycoscience and Biocatalysis, Department of Bio-Organic Chemistry, Bijvoet Center, Utrecht University, The Netherlands.
Dudziak
Gregor
G
Kragl
Udo
U
Wandrey
Christian
C
Kamerling
Johannis P
JP
Vliegenthart
Johannes F G
JF
D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D019906:Nuclear Magnetic Resonance, Biomolecular; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D011134:Polysaccharides / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D000080506:Sialyl Lewis X Antigen; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D018396:Mucin-1; D009844:Oligosaccharides; D011134:Polysaccharides; D000080506:Sialyl Lewis X Antigen