Automated Solid‐Phase Synthesis of Chondroitin Sulfate Glycosaminoglycans

Eller, Steffen; Collot, Mayeul; Yin, Jian; Hahm, Heung Sik; Seeberger, Peter H.

doi:10.1002/anie.201210132

Cited by 157 publications

(187 citation statements)

References 35 publications

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“…All automated glycosylations were performed on an automated oligosaccharide synthesizer demonstrator unit using anhydrous solvents of the cycle-Tainer solvent delivery system. Oligosaccharides 1-9 were synthesized following the schemes in Extended Data Figs 1 and 2 using sequences I to V (Extended Data Table 1) 11,27,[31][32][33] . Detailed synthesis information and NMR data can be obtained from the Supplementary Information.…”

Section: Methodsmentioning

confidence: 99%

Identification of carbohydrate anomers using ion mobility–mass spectrometry

Hofmann

Hahm

Seeberger

et al. 2015

Nature

299

297

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Carbohydrates are ubiquitous biological polymers that are important in a broad range of biological processes. However, owing to their branched structures and the presence of stereogenic centres at each glycosidic linkage between monomers, carbohydrates are harder to characterize than are peptides and oligonucleotides. Methods such as nuclear magnetic resonance spectroscopy can be used to characterize glycosidic linkages, but this technique requires milligram amounts of material and cannot detect small amounts of coexisting isomers. Mass spectrometry, on the other hand, can provide information on carbohydrate composition and connectivity for even small amounts of sample, but it cannot be used to distinguish between stereoisomers. Here, we demonstrate that ion mobility-mass spectrometry--a method that separates molecules according to their mass, charge, size, and shape--can unambiguously identify carbohydrate linkage-isomers and stereoisomers. We analysed six synthetic carbohydrate isomers that differ in composition, connectivity, or configuration. Our data show that coexisting carbohydrate isomers can be identified, and relative concentrations of the minor isomer as low as 0.1 per cent can be detected. In addition, the analysis is rapid, and requires no derivatization and only small amounts of sample. These results indicate that ion mobility-mass spectrometry is an effective tool for the analysis of complex carbohydrates. This method could have an impact on the field of carbohydrate synthesis similar to that of the advent of high-performance liquid chromatography on the field of peptide assembly in the late 1970s.

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Section: Methodsmentioning

confidence: 99%

Identification of carbohydrate anomers using ion mobility–mass spectrometry

Hofmann

Hahm

Seeberger

et al. 2015

Nature

299

297

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“…Although the synthesis of CS oligosaccharides has been studied extensively, [8][9][10][11][12][13][14][15][16][17][18] the design and synthesis of the building blocks for generating a CS library is still challenging. In the present study, to exhaustively investigate the binding properties of proteins or viruses at the molecular level, we demonstrated the systematic synthesis of all 16 kinds of CS partial disaccharide structures possessing a diverse sulfation set.…”

mentioning

confidence: 99%

Synthesis of a chondroitin sulfate disaccharide library and a GAG-binding protein interaction analysis

Wakao¹,

Obata²,

Miyachi³

et al. 2015

Bioorganic & Medicinal Chemistry Letters

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Chondroitin sulfate (CS), which belongs to the glycosaminoglycan (GAG) superfamily, is a linear sulfated polysaccharide involved in various biological processes. CS structure is very heterogeneous and contains various sulfation patterns owing to the multiple and random enzymatic modifications that occur during its biosynthesis. The resultant microdomain structure in the CS chain interacts with specific biomolecules to regulate biological functions. Therefore, an analysis of the structure-activity relationship of CS at the molecular level is necessary to clarify their biofunctions. In this study, we designed the common intermediate possessing an orthogonally removable protective group and systematically synthesized all 16 types of CS disaccharide structure generated by sulfation. In addition, we demonstrated the on-time analysis of the binding properties of GAG-binding proteins using 'Sugar Chip' immobilized CS disaccharide structures by surface plasmon resonance (SPR) imaging, indicating that our chip technology is effective for the evaluation of binding properties.

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“…Following selective removal of the Lev group at the branching position without cleavage of the Fmoc group at the non-reducing end glucose residue, the b-(1,6)-branch was introduced using module A1, and the two remaining Fmoc groups were removed. Following cleavage of the crude product from the resin by UV irradiation in flow, 29 HPLC analysis showed one major peak (Fig. 2).…”

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confidence: 99%

“…Merrifield resin functionalized with photolabile linker 4 served as solid support for the incorporation of the monosaccharide building blocks. 29 To avoid problems caused by residual base remaining after deprotection, 27 an acidic washing step (TMSOTf in CH 2 Cl 2 at À30 1C for 1 min) was introduced, preceding each glycosylation cycle and ensuring the removal of residual base and water after the previous deprotection step. The assembly of branched b-(1,3)-glucan pentasaccharide 5 served as a test for our automated glycan assembly (AGA) approach (Scheme 1).…”

mentioning

confidence: 99%