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Chemical Synthesis and Enzymatic Testing of CMP-Sialic Acid Derivatives

Authors

  • Dr. Saskia Wolf,

    1. Organic Chemistry, Department of Chemistry, Faculty of Sciences, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany)
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  • Dr. Svenja Warnecke,

    1. Organic Chemistry, Department of Chemistry, Faculty of Sciences, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany)
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  • Jörg Ehrit,

    1. Institut für Zelluläre Chemie, Medizinische Hochschule Hannover, Carl-Neuberg-Straße 1, 30625 Hannover (Germany)
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  • Dr. Friedrich Freiberger,

    1. Institut für Zelluläre Chemie, Medizinische Hochschule Hannover, Carl-Neuberg-Straße 1, 30625 Hannover (Germany)
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  • Prof. Dr. Rita Gerardy-Schahn,

    1. Institut für Zelluläre Chemie, Medizinische Hochschule Hannover, Carl-Neuberg-Straße 1, 30625 Hannover (Germany)
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  • Prof. Dr. Chris Meier

    Corresponding author
    1. Organic Chemistry, Department of Chemistry, Faculty of Sciences, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany)
    • Organic Chemistry, Department of Chemistry, Faculty of Sciences, University of Hamburg, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany)
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Abstract

The cycloSal approach has been used in the past for the synthesis of a range of phosphorylated bioconjugates. In those reports, cycloSal nucleotides were allowed to react with different phosphate nucleophiles. With glycopyranosyl phosphates as nucleophiles, diphosphate-linked sugar nucleotides were formed. Here, cycloSal-nucleotides were used to prepare monophosphate-linked sugar nucleotides successfully in high anomeric purity and high chemical yield. The method was successfully used for the synthesis of three nucleotide glycopyranoses as model compounds. The method was then applied to the syntheses of CMP-N-acetyl-neuraminic acids (CMP-Neu5NAc) and of four derivatives with different modifications at their amino functions (N-propanoyl, N-butanoyl, N-pentanoyl and N-cyclopropylcarbonyl). The compounds were used for initial enzymatic studies with a bacterial polysialyltransferase (polyST). Surprisingly, the enzyme showed marked differences in terms of utilisation of the four derivatives. The N-propanoyl, N-butanoyl, and N-pentanoyl derivatives were efficiently used in a first transfer with a fluorescently labelled trisialo-acceptor. However, elongation of the resulting tetrasialo-acceptors worsened progressively with the size of the N-acyl chain. The N-pentanoyl derivative allowed a single transfer, leading to a capped tetramer. The N-cyclopropylcarbonyl derivative was not transferred.

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