Polysialic acid engineering: synthesis of polysialylated neoglycosphingolipids by using the polysialyltransferase from neuroinvasive Escherichia coli K1.

Cho, Jin Won; Troy, Frederic A.

doi:10.1073/pnas.91.24.11427

Cited by 41 publications

(20 citation statements)

References 39 publications

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“…The requirement in PSA biosynthesis for an initiator of some sort derives from the inability of NeuS to synthesize PSA when presented with exogenous CMP-Sia (133). In contrast, NeuS will elongate (albeit with weak processivity) oligosialic acid acceptors containing three or more sialic acid residues (131) and will efficiently elongate certain ganglioside (sialylated mammalian glycolipid) acceptors in vitro (27,85). However, the fact remains that exogenous acceptors must already contain sialic acid, which does not help explain how at least the first sialic acid residue is added to the (unknown) endogenous acceptors in E. coli K1 PSA biosynthesis.…”

Section: De Novo Synthesismentioning

confidence: 99%

Diversity of Microbial Sialic Acid Metabolism

Vimr

Kalivoda

Deszo

et al. 2004

Microbiol Mol Biol Rev

516

663

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Sialic acids are structurally unique nine-carbon keto sugars occupying the interface between the host and commensal or pathogenic microorganisms. An important function of host sialic acid is to regulate innate immunity, and microbes have evolved various strategies for subverting this process by decorating their surfaces with sialylated oligosaccharides that mimic those of the host. These subversive strategies include a de novo synthetic pathway and at least two truncated pathways that depend on scavenging host-derived intermediates. A fourth strategy involves modification of sialidases so that instead of transferring sialic acid to water (hydrolysis), a second active site is created for binding alternative acceptors. Sialic acids also are excellent sources of carbon, nitrogen, energy, and precursors of cell wall biosynthesis. The catabolic strategies for exploiting host sialic acids as nutritional sources are as diverse as the biosynthetic mechanisms, including examples of horizontal gene transfer and multiple transport systems. Finally, as compounds coating the surfaces of virtually every vertebrate cell, sialic acids provide information about the host environment that, at least in Escherichia coli, is interpreted by the global regulator encoded by nanR. In addition to regulating the catabolism of sialic acids through the nan operon, NanR controls at least two other operons of unknown function and appears to participate in the regulation of type 1 fimbrial phase variation. Sialic acid is, therefore, a host molecule to be copied (molecular mimicry), eaten (nutrition), and interpreted (cell signaling) by diverse metabolic machinery in all major groups of mammalian pathogens and commensals

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Section: De Novo Synthesismentioning

confidence: 99%

Diversity of Microbial Sialic Acid Metabolism

Vimr

Kalivoda

Deszo

et al. 2004

Microbiol Mol Biol Rev

516

663

View full text Add to dashboard Cite

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“…We concluded that all bacterial polySTs possess structurally similar acceptor binding sites. However, exogenous sialyl oligomers are poorly elongated (23)(24)(25), with most acting as acceptor for a single sialyl addition unless tethered to the membrane by a lipid anchor (26). Therefore, broadly extrapolating results derived from the use of exogenous acceptors to natural PSA elongation may be suspect to misinterpretation because of the failure to duplicate the in vivo process.…”

Section: Exogenous Acceptor Activities Of Heterologously Expressedmentioning

confidence: 99%

Functional Relationships of the Sialyltransferases Involved in Expression of the Polysialic Acid Capsules ofEscherichia coli K1 and K92 and Neisseria meningitidis Groups B or C

Steenbergen

Vimr

2003

Journal of Biological Chemistry

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Polysialic acid (PSA) capsules are cell-associated homopolymers of ␣2,8-, ␣2,9-, or alternating ␣2,8/2,9-linked sialic acid residues that function as essential virulence factors in neuroinvasive diseases caused by certain strains of Escherichia coli and Neisseria meningitidis. PSA chains structurally identical to the bacterial ␣2,8-linked capsular polysaccharides are also synthesized by the mammalian central nervous system, where they regulate neuronal function in association with the neural cell adhesion molecule (NCAM). Despite the structural identity between bacterial and NCAM PSAs, the respective polysialyltransferases (polySTs) responsible for polymerizing sialyl residues from donor CMP-sialic acid are not homologous glycosyltransferases. To better define the mechanism of capsule biosynthesis, we established the functional interchangeability of bacterial polySTs by complementation of a polymerase-deficient E. coli K1 mutant with the polyST genes from groups B or C N. meningitidis and the control E. coli K92 polymerase gene. The biochemical and immunochemical results demonstrated that linkage specificity is dictated solely by the source of the polymerase structural gene. To determine the molecular basis for linkage specificity, we created chimeras of the K1 and K92 polySTs by overlap extension PCR. Exchanging the first 52 N-terminal amino acids of the K1 NeuS with the C terminus of the K92 homologue did not alter specificity of the resulting chimera, whereas exchanging the first 85 or reciprocally exchanging the first 100 residues did. These results demonstrated that linkage specificity is dependent on residues located between positions 53 and 85 from the N terminus. Site-directed mutagenesis of the K92 polyST N terminus indicated that no single residue alteration was sufficient to affect specificity, consistent with the proposed function of this domain in orienting the acceptor. The combined results provide the first evidence for residues critical to acceptor binding and elongation in polysialyltransferase.

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“…The a-2,6-sialyltransferase from Photobacterium damsela has been shown to transfer sialic acid to terminal galactose residues that are fucosylated or sialylated at the 2 or 3 position, respectively [I I], and such an acceptor specificity has not been reported for mammalian sialyltransferases. The a-2,8-sialyltransferase from E. coli has been used in the synthesis of glycosphingolipids [12], while crude extracts of E. coli expressing the N. meningitidis a-2,3-sialyltransferase have been used to sialylate a lactose derivative [ 131. The difference in their specificities makes these bacterial transferases important tools for the chemienzymatic synthesis of oligosaccharides with biological activity.…”

mentioning

confidence: 99%

Characterization of a Recombinant Neisseria Meningitidesα‐2,3‐Sialyltransferase and its Acceptor Specificity

Gilbert

Cunningham

Watson

et al. 1997

European Journal of Biochemistry

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The structure and specificity of the recombinant a-2,3-sialyltransferase from Neisseria meningitidis are reported. This enzyme showed an unusual acceptor specificity in that it could use a-terminal and pterminal Gal residues as acceptors. In addition (pl-+4)-linked and (pl+3)-linked terminal Gal served as acceptors. These properties distinguish the bacterial enzyme from the more widely investigated mammalian equivalents. The protein was expressed as a membrane-associated protein in Escherichia coli at a level of 750 U/I (-250 mgfl). The protein could be extracted with buffers containing 0.2% Triton X-100 and purified to homogeneity using immobilized-metal-affinity chromatography. Electrospray-ionization mass spectrometry of peptides obtained by cleavage with cyanogen bromide and trypsin confirmed over 95 % of the deduced amino acid sequence. When used for enzymatic synthesis in coupled reactions with recombinant CMP-NeuSAc synthetase, the a-2,3-sialyltransferase could sialylate fluorescent derivatives of N-acetyllactosamine with N-acetylneuraminic acid, N-propionylneuraminic acid and N-glycoloylneuraminic acid.Keywords: Neisseria meningitidis ; sialyltransferase Sialyltransferases are a group of glycosyltransferases that transfer sialic acid (N-acetylneuraminic acid, NeuSAc) from the activated sugar nucleotide CMP-NeuSAc to acceptor oligosaccharides found on glycoproteins, glycolipids or polysaccharides. It has become apparent that these sialylated oligosaccharides play important roles in cell-cell recognition, cell differentiation, and various receptor-ligand interactions in mammalian systems. The large number of sialylated oligosaccharide structures has led to the characterization of many sialyltransferases involved in the synthesis of these structures. Based on the linkage and acceptor specificity of the sialyltransferases studied, it has been determined that at least 13 sialyltransferase genes are present in mammalian systems [l].Sialylated glycoconjugates are also found in bacteria [2-31, where they are structures that mimic oligosaccharides found in mammalian glycolipids [4] and are probably used to evade the host immune response. The importance of sialylated lipo-oligosaccharides (LOS) in the pathogenesis of Neisseria gonorrhoeae has been established [5], while for Neisseria meningitidis the polysialic acid capsule and the sialylated LOS were found to be important for pathogenicity [6]. Despite their importance as proven or potential virulence factors, few bacterial sialyltransferases have been cloned [7-91 or purified [lo]. The a-2,8-sialyltransferases involved in the synthesis of the polysialic acid capsules have been cloned and expressed from both Escherichia coli [7] and N. meningitidis [S], and we have recently cloned the LOS a-2,3-sialyltransferases from N. meningitidis and N. gonorrhoeae [9].Because of the biological activity of their products, mammalian sialyltransferases are expected to act in specific tissues, cell compartments and/or developmental stages to create precise sialyloglycans. Bact...

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Polysialic acid engineering: synthesis of polysialylated neoglycosphingolipids by using the polysialyltransferase from neuroinvasive Escherichia coli K1.

Cited by 41 publications

References 39 publications

Diversity of Microbial Sialic Acid Metabolism

Diversity of Microbial Sialic Acid Metabolism

Functional Relationships of the Sialyltransferases Involved in Expression of the Polysialic Acid Capsules ofEscherichia coli K1 and K92 and Neisseria meningitidis Groups B or C

Characterization of a Recombinant Neisseria Meningitidesα‐2,3‐Sialyltransferase and its Acceptor Specificity

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