Chemoenzymatic Synthesis of a MUC1 Glycopeptide Carrying Non-Natural Sialyl TF-β O-Glycan

A MUC1 type of glycopeptide was synthesized by the 9-fluorenylmethoxycarbonyl (Fmoc) solid-phase peptide synthesis (SPPS) protocol using benzyl and benzylidene-protected β-D-Gal-(1→3)-β-D-GalNAc-Ser/Thr (TF-β: a stereoisomer of the Thomsen-Friedenreich antigen). The synthetic glycopeptide was released from the resin with reagent K, and the resulting benzylated glycopeptide was deprotected under conditions of low-acidity trifluoromethanesulfonic acid (TfOH). The glycopeptide carrying duplicate non-natural O-glycans was dominant in the products, but was accompanied by a considerable amount of the glycopeptide missing one of the O-glycans. In contrast, the native α-glycoside was relatively stable under the acidic debenzylation conditions as shown by a parallel experiment with the glycopeptide involving α-D-GalNAc-Ser/Thr linkage. Enzymatic glycosylation with CMP-NeuAc was successful with both natural and non-natural O-glycans of the synthetic glycopeptide.

[1]  A. Suzuki,et al.  Chemical synthesis of 23 kDa glycoprotein by repetitive segment condensation: a synthesis of MUC2 basal motif carrying multiple O-GalNAc moieties. , 2005, Journal of the American Chemical Society.

[2]  S. Nishimura,et al.  Combinatorial synthesis of MUC1 glycopeptides: polymer blotting facilitates chemical and enzymatic synthesis of highly complicated mucin glycopeptides. , 2005, Journal of the American Chemical Society.

[3]  M. Hollingsworth,et al.  Thomsen-Friedenreich antigen expression in gastric carcinomas is associated with MUC1 mucin VNTR polymorphism. , 2005, Glycobiology.

[4]  S. Nishimura,et al.  Rapid microwave-assisted solid-phase glycopeptide synthesis. , 2005, Organic letters.

[5]  H. Hojo,et al.  Solid-Phase Synthesis of Core 8 O-Glycan-Linked MUC5AC Glycopeptide , 2005, Bioscience, biotechnology, and biochemistry.

[6]  U. Karsten,et al.  Binding patterns of DTR-specific antibodies reveal a glycosylation-conditioned tumor-specific epitope of the epithelial mucin (MUC1). , 2004, Glycobiology.

[7]  H. Hojo,et al.  Synthesis of a mimic for the heterogeneous surface of core 2 sialoglycan-linked glycoprotein. , 2004, Organic letters.

[8]  D. Kufe,et al.  Human MUC1 carcinoma-associated protein confers resistance to genotoxic anticancer agents. , 2004, Cancer cell.

[9]  Yukishige Ito,et al.  Synthesis of N-linked pentasaccharides with isomeric glycosidic linkage , 2000, Glycoconjugate Journal.

[10]  H. Hojo,et al.  Solid-phase synthesis of core 2 O-linked glycopeptide and its enzymatic sialylation , 2003 .

[11]  H. Kunz,et al.  Synthesis of a Tumor-Associated2,3-Sialyl-T Glycododecapeptide Antigen from the Tandem Repeat Regionof the Mucin MUC1 , 2003 .

[12]  H. Hojo,et al.  Synthesis of an Unnatural N-Glycan-linked Dolichyl Pyrophosphate Precursor , 2003, Bioscience, biotechnology, and biochemistry.

[13]  M. Someya,et al.  Preparation of core 2 type tetrasaccharide carrying decapeptide by benzyl protection-based solid-phase synthesis strategy , 2002 .

[14]  A. Liese,et al.  Chemoenzymatic-Chemical Synthesis of a (2-3)-Sialyl T Threonine Building Block and Its Application to the Synthesis of the N-Terminal Sequence of Leukemia-Associated Leukosialin (CD 43). , 2001, Angewandte Chemie.

[15]  Y. Ito,et al.  Solid-phase synthesis of the glycopeptide of human glycophorin AM bearing the consecutive sialyl-T antigen. , 2000, Carbohydrate Research.

[16]  S. Müller,et al.  MUC1: the polymorphic appearance of a human mucin. , 2000, Glycobiology.

[17]  J. Taylor‐Papadimitriou,et al.  MUC1 and cancer. , 1999, Biochimica et biophysica acta.

[18]  Yukishige Ito,et al.  A novel silyl linker : Motif for side chain tethered approach to solid-phase glycopeptide synthesis , 1999 .

[19]  Yukishige Ito,et al.  Total synthesis of B-chain of human α2HS glycoprotein , 1997 .

[20]  Y. Nakahara,et al.  Solid-phase synthesis of an O-linked glycopeptide based on a benzyl-protected glycan approach. , 1996, Carbohydrate research.

[21]  J. Peter-Katalinic,et al.  MUC1 Glycoforms in Breast Cancer , 1996 .

[22]  J. Peter-Katalinic,et al.  MUC1 glycoforms in breast cancer--cell line T47D as a model for carcinoma-associated alterations of 0-glycosylation. , 1996, European journal of biochemistry.

[23]  I. Brockhausen,et al.  Mechanisms underlying aberrant glycosylation of MUC1 mucin in breast cancer cells. , 1995, European journal of biochemistry.

[24]  F. Hanisch,et al.  Monoclonal antibody BW494 defines a blood group Lewisa/type 1 chain-related antigen on carcinoma-associated mucins. , 1993, Biological chemistry Hoppe-Seyler.