Functional neoglycopeptides

synthesis and characterization of a new class of MUC1 glycoprotein models having core 2-based O-glycan and complex-type N-glycan chains

Takahiko Matsushita, Reiko Sadamoto, Naoki Ohyabu, Hideki Nakata, Masataka Fumoto, Naoki Fujitani, Yasuhiro Takegawa, Takeshi Sakamoto, Masaki Kurogochi, Hiroshi Hinou, Hiroki Shimizu, Takaomi Ito, Kentarou Naruchi, Hiroko Togame, Hiroshi Takemoto, Hirosato Kondo, Shin Ichiro Nishimura

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

An efficient protocol for the construction of MUC1-related glycopeptide analogues having complex O-glycan and N-glycan chains was established by integrating chemical and enzymatic approaches on the functional polymer platforms. We demonstrated the feasibility of sortase A-mediated ligation between two glycopeptide segments by tagging with signal peptides, LPKTGLR and GG, at each C- or N-terminal position. Structural analysis of the macromolecular N,O-glycopeptides was performed by means of ESI-TOFMS (MS/MS) equipped with an electron-captured dissociation device. Immunological assay using MUC1 glycopeptides synthesized in this study revealed that N-glycosylation near the antigenic O-glycosylated PDTR motif did not disturb the interaction between the anti-MUC1 monoclonal antibody and this crucial O-glycopeptide moiety.NMR study indicated that the N-terminal immunodominant region [Ala-Pro-Asp-Thr(O-glycan)- Arg] forms an inverse γ-turn-like structure, while the C-terminal region composed of N-glycopeptide and linker SrtA-peptide was proved to be an independently random structure. These results indicate that the bulky O- and N-glycan chains can function independently as disease-relevant epitopes and ligands for carbohydrate-binding proteins, when both are combined by an artificial intervening peptide having a possible effect of separating N- and C-terminal regions. The present strategy will greatly facilitate rapid synthesis of multiply functionalized complex neoglycopeptides as new types of convenient tools or models for the investigation of thhe structure-function relationship of various glycoproteins and development of novel class glycopeptide-based biopharmaceuticals, drug delivery systems, and biomedical materials.

Original languageEnglish
Pages (from-to)11117-11133
Number of pages17
JournalBiochemistry
Volume48
Issue number46
DOIs
Publication statusPublished - Nov 24 2009
Externally publishedYes

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Glycopeptides
Polysaccharides
Glycoproteins
Glycosylation
Immunodominant Epitopes
Functional polymers
Peptides
Drug Delivery Systems
Protein Sorting Signals
Structural analysis
Ligation
Epitopes
Assays
Polymers
Monoclonal Antibodies
Nuclear magnetic resonance
Electrons
Ligands
Equipment and Supplies

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Functional neoglycopeptides : synthesis and characterization of a new class of MUC1 glycoprotein models having core 2-based O-glycan and complex-type N-glycan chains. / Matsushita, Takahiko; Sadamoto, Reiko; Ohyabu, Naoki; Nakata, Hideki; Fumoto, Masataka; Fujitani, Naoki; Takegawa, Yasuhiro; Sakamoto, Takeshi; Kurogochi, Masaki; Hinou, Hiroshi; Shimizu, Hiroki; Ito, Takaomi; Naruchi, Kentarou; Togame, Hiroko; Takemoto, Hiroshi; Kondo, Hirosato; Nishimura, Shin Ichiro.

In: Biochemistry, Vol. 48, No. 46, 24.11.2009, p. 11117-11133.

Research output: Contribution to journalArticle

Matsushita, T, Sadamoto, R, Ohyabu, N, Nakata, H, Fumoto, M, Fujitani, N, Takegawa, Y, Sakamoto, T, Kurogochi, M, Hinou, H, Shimizu, H, Ito, T, Naruchi, K, Togame, H, Takemoto, H, Kondo, H & Nishimura, SI 2009, 'Functional neoglycopeptides: synthesis and characterization of a new class of MUC1 glycoprotein models having core 2-based O-glycan and complex-type N-glycan chains', Biochemistry, vol. 48, no. 46, pp. 11117-11133. https://doi.org/10.1021/bi901557a
Matsushita, Takahiko ; Sadamoto, Reiko ; Ohyabu, Naoki ; Nakata, Hideki ; Fumoto, Masataka ; Fujitani, Naoki ; Takegawa, Yasuhiro ; Sakamoto, Takeshi ; Kurogochi, Masaki ; Hinou, Hiroshi ; Shimizu, Hiroki ; Ito, Takaomi ; Naruchi, Kentarou ; Togame, Hiroko ; Takemoto, Hiroshi ; Kondo, Hirosato ; Nishimura, Shin Ichiro. / Functional neoglycopeptides : synthesis and characterization of a new class of MUC1 glycoprotein models having core 2-based O-glycan and complex-type N-glycan chains. In: Biochemistry. 2009 ; Vol. 48, No. 46. pp. 11117-11133.
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abstract = "An efficient protocol for the construction of MUC1-related glycopeptide analogues having complex O-glycan and N-glycan chains was established by integrating chemical and enzymatic approaches on the functional polymer platforms. We demonstrated the feasibility of sortase A-mediated ligation between two glycopeptide segments by tagging with signal peptides, LPKTGLR and GG, at each C- or N-terminal position. Structural analysis of the macromolecular N,O-glycopeptides was performed by means of ESI-TOFMS (MS/MS) equipped with an electron-captured dissociation device. Immunological assay using MUC1 glycopeptides synthesized in this study revealed that N-glycosylation near the antigenic O-glycosylated PDTR motif did not disturb the interaction between the anti-MUC1 monoclonal antibody and this crucial O-glycopeptide moiety.NMR study indicated that the N-terminal immunodominant region [Ala-Pro-Asp-Thr(O-glycan)- Arg] forms an inverse γ-turn-like structure, while the C-terminal region composed of N-glycopeptide and linker SrtA-peptide was proved to be an independently random structure. These results indicate that the bulky O- and N-glycan chains can function independently as disease-relevant epitopes and ligands for carbohydrate-binding proteins, when both are combined by an artificial intervening peptide having a possible effect of separating N- and C-terminal regions. The present strategy will greatly facilitate rapid synthesis of multiply functionalized complex neoglycopeptides as new types of convenient tools or models for the investigation of thhe structure-function relationship of various glycoproteins and development of novel class glycopeptide-based biopharmaceuticals, drug delivery systems, and biomedical materials.",
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AU - Ohyabu, Naoki

AU - Nakata, Hideki

AU - Fumoto, Masataka

AU - Fujitani, Naoki

AU - Takegawa, Yasuhiro

AU - Sakamoto, Takeshi

AU - Kurogochi, Masaki

AU - Hinou, Hiroshi

AU - Shimizu, Hiroki

AU - Ito, Takaomi

AU - Naruchi, Kentarou

AU - Togame, Hiroko

AU - Takemoto, Hiroshi

AU - Kondo, Hirosato

AU - Nishimura, Shin Ichiro

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