Chemoenzymatic synthesis of sialylated glycopeptides derived from mucins and T-cell stimulating peptides.
The Tn, T, sialyl-Tn, and 2,3-sialyl-T antigens are tumor-associated carbohydrate antigens expressed on mucins in epithelial cancers, such as those affecting the breast, ovary, stomach, and colon. Glycopeptides carrying these antigens are of interest for development of cancer vaccines and a short, chemoenzymatic strategy for their synthesis is reported. Building blocks corresponding to the Tn (GalNAc alpha-Ser/Thr) and T [Gal beta(1-->3)GalNAc alpha-Ser/Thr] antigens, which are relatively easy to obtain by chemical synthesis, were prepared and then used in the synthesis of glycopeptides on the solid phase. Introduction of sialic acid to give the sialyl-Tn [Neu5Ac alpha(2-->6)GalNAc alpha-Ser/Thr] and 2,3-sialyl-T [Neu5Ac alpha(2-->3)Gal beta(1-->3)GalNAc alpha-Ser/Thr] antigens is difficult when performed chemically at the building block level. Sialylation was therefore carried out with recombinant sialyltransferases in solution after cleavage of the Tn and T glycopeptides from the solid phase. In the same manner, the core 2 trisaccharide [Gal beta 1-->3(GlcNAc beta 1-->6)GalNAc] was incorporated in glycopeptides containing the T antigen by using a recombinant N-acetylglucosaminyltransferase. The outlined chemoenzymatic approach was applied to glycopeptides from the tandem repeat domain of the mucin MUC1, as well as to neoglycosylated derivatives of a T cell stimulating viral peptide.
George
S K
SK
Department of Organic Chemistry, Umeå University, SE-901 87 Umeå, Sweden.
Schwientek
T
T
Holm
B
B
Reis
C A
CA
Clausen
H
H
Kihlberg
J
J
D000595:Amino Acid Sequence; D000818:Animals; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000737:chemistry*; D002240:Carbohydrate Sequence; D018984:Epitopes, T-Lymphocyte / Q000737:chemistry; D006020:Glycopeptides / Q000138:chemical synthesis*; D006801:Humans; D007313:Insecta; D051379:Mice; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000737:chemistry*; D009077:Mucins / Q000737:chemistry; D017351:N-Acetylglucosaminyltransferases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry* / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D012799:Sialyltransferases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects* / Q000276:immunology
D015295:Antigens, Tumor-Associated, Carbohydrate; D018984:Epitopes, T-Lymphocyte; D006020:Glycopeptides; D018396:Mucin-1; D009077:Mucins; D010455:Peptides; D011994:Recombinant Proteins; C040286:Tn antigen; C055491:sialosyl-Tn antigen; C016467:Thomsen-Friedenreich antigen; D017351:N-Acetylglucosaminyltransferases; C053024:beta-1,3-galactosyl-O-glycosyl-glycoprotein beta-1,6-acetylglucosaminyl transferase; D012799:Sialyltransferases; C038525:CMP-N-acetylneuraminate-alpha-N-acetylgalactosaminide alpha-2,6-sialyltransferase; D019158:N-Acetylneuraminic Acid