Construction and structural characterization of versatile lactosaminoglycan-related compound library for the synthesis of complex glycopeptides and glycosphingolipids

Kentarou Naruchi, Tomoki Hamamoto, Masaki Kurogochi, Hiroshi Hinou, Hiroki Shimizu, Takahiko Matsushita, Naoki Fujitani, Hirosato Kondo, Shin Ichiro Nishimura

Research output: Contribution to journalArticlepeer-review

37 Citations (Scopus)

Abstract

We have established a facile and efficient protocol for the preparative-scale synthesis of various compound libraries related to lactosaminoglycans: cell surface oligosaccharides composed of N-acetyllactosamine as a repeating disaccharide unit, based on chemical and enzymatic approaches. Substrate specificity and feasibility of a bacterial glycosyltransferase, Neisseria meningitidis β1,3-N- acetylglucosaminyltransferase (LgtA), were investigated in order to synthesize various key intermediates suited for the construction of mammalian O-glycopeptides and glycosphingolipids containing poly-N-acetyllactosamine structures. Recombinant LgtA exhibited the highest glycosyltransferase activity with strongly basic conditions (pH = 10, glycine-NaOH buffer) and a broad range of optimal temperatures from 20 to 30 °C. Interestingly, it was found that LgtA discriminates L-serine and L-threonine and functions both as a core-1 β1,3-N-acetylglucosaminyltransferase and core-2 β1,3-N- acetylglucosaminyltransferase toward Fmoc-Ser derivatives, while LgtA showed only core-2 β1,3-N-acetylglucosaminyltransferase activity in the presence of Fmoc-Thr derivatives. Combined use of LgtA with human β1,4- galactosyltransferase allowed for controlled sugar extension reactions from synthetic sugar amino acids and gave synthetic lactosaminoglycans, such as a decasaccharide derivative, Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 3)]GalNAcα1 → Fmoc-Ser-OH (6), and a dodecasaccharide derivative, Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 3)Galβ(1 → 3)]GalNAcα1 → Fmoc-Ser-OH (9). A partially protected pentasaccharide intermediate, GlcNAcβ(1 → 3)Galβ(1 → 4)GlcNAcβ(1 → 6)[Galβ(1 → 3)]GalNAcα1 → Fmoc-Thr-OH (11), was applied for the microwave-assisted solid-phase synthesis of a MUC1-related glycopeptide 19 (MW = 2610.1). The findings suggest that this sugar extension strategy can be employed for the modification of lactosyl ceramide mimetic polymers to afford convenient precursors for the synthesis of various glycosphingolipids.

Original languageEnglish
Pages (from-to)9609-9621
Number of pages13
JournalJournal of Organic Chemistry
Volume71
Issue number26
DOIs
Publication statusPublished - Dec 22 2006
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Organic Chemistry

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