Enzyme-catalyzed synthesis of carbohydrates

Toone, Eric J.; Simon, Ethan S.; Bednarski, Mark D.; Whitesides, George M.

doi:10.1016/s0040-4020(01)89487-4

Cited by 374 publications

(68 citation statements)

References 217 publications

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“…broad use in the study of carbohydrate binding by antibodies, The use of enzyme assisted oligosaccharide synthesis is gaining wide popularity since it allows the rapid and specific homologation of relatively small, readily chemically synthesized mono-and disaccharides into larger and morecomplex structures (1). This homologation is most frequently accomplished using specific glycosyltransferases that transfer sugars from the appropriate sugar-nucleotide donors, all of which are commercially available, to suitable oligosaccharide acceptors.…”

Section: Introductionmentioning

confidence: 99%

“…We present here an investigation using a readily isolated Lewis blood-group associated a(1-4) fucosyltransferase, a glycosyltransferase that has previously been used in the combined chemical-enzymatic synthesis of oligosaccharides terminating in the Lewis-a (Lea) blood-group determinant (5). The Lewis fucosyltransferase (Le-Fuc-T), which has also recently been cloned (2), transfers an L-fucopyranosyl residue from guanosine-5'-diphospho-fucose (GDP-fucose, 5) to the 4-hydroxyl group of oligosaccharides terminating in the Type 1 sequence P D G~~(~-~) P D G~C N A C -O R (I), to produce the Lea-active terminus @Gal (1)(2)(3) [ a~F u c ( 1-4:1] PcGlcNAc-OR (2) (5). This reaction has been kinetically characterized in vitro where R in the synthetic oligosaccharides 1 and 2 was the 8-methoxycarbonyloctyl aglycone (5).…”

Section: Introductionmentioning

confidence: 99%

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Chemical synthesis of GDP-fucose analogs and their utilization by the Lewis *A(1 → 4) fucosyltransferase

Gokhale¹,

Hindsgaul²,

Palcic³

1990

Can. J. Chem.

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Chemical syntheses are reported for GDP-fucose (S), GDP-3-deoxy-fucose (6), and GDP-arabinose (7), the demethyl analog of 5. All three sugar nucleotides were found to act as donor substrates for an a(1+4) fucosyltransferase isolated from human milk when PDG~~(~+~)PDG~~NAC-O(CH~)~COOM~ (1) was used as the acceptor. The rate of transfer of sugar residues to 1 was measured using a coupled spectrophotometric assay and was found to be 100% (S), 2.3% (6), and 5.9% (7). The product Lea-active oligosaccharide analogs were identified by both an enzyme-linked immunosorbent assay (ELISA) and by 'H NMR spectroscopy.Key words: glycosyltransferase, oligosaccharide synthesis, sugar-nucleotide analog, ELISA assay, fucosyltransferase.UDAY B. GOKHALE, OLE HINDSGAUL et MONICA M. PALCIC. Can. J. Chem. 68, 1063(1990.On a rCalisC la synthkse du GDP-fucose (S), du GDP-3-desoxy-fuxose (6) et du GDP-arabinose (7) Introduction broad use in the study of carbohydrate binding by antibodies, The use of enzyme assisted oligosaccharide synthesis is gaining wide popularity since it allows the rapid and specific homologation of relatively small, readily chemically synthesized mono-and disaccharides into larger and morecomplex structures (1). This homologation is most frequently accomplished using specific glycosyltransferases that transfer sugars from the appropriate sugar-nucleotide donors, all of which are commercially available, to suitable oligosaccharide acceptors. Accessibility to the required mammalian glycosyltransferases, most of which must still be isolated from natural sources, has been the single major impediment to the general applicability of this combined chemical-enzymatic approach to oligosaccharide synthesis. The availability of the required enzymes is, however, rapidly increasing as three of these enzymes are now commercially available and at least six have recently been cloned (2).Analogs of the naturally occumng oligosaccharides, particularly deoxygenated oligosaccharides, are invaluable probes in studies on the molecular recognition of complex carbohydrates by protein combining sites (3, 4). The effect of specifically deoxygenating individual hydroxyl groups of oligosaccharide ligands, for example, provides direct evidence for the involvement of those hydroxyl groups in hydrogen bonding with the protein. While the utility of oligosaccharide analogs is generally recognized, the difficulty of preparing the required structures by multistep chemical synthesis remains a major obstacle to their

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Chemical synthesis of GDP-fucose analogs and their utilization by the Lewis *A(1 → 4) fucosyltransferase

Gokhale¹,

Hindsgaul²,

Palcic³

1990

Can. J. Chem.

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“…Chemical synthesis of oligosaccharides requires sophisticated strategies for protecting/deprotecting and assembling sugar monomers and for controlling product regiochemistry and stereochemistry (8)(9)(10). Enzymatic synthesis using glycosyltransferases has emerged as a useful alternative to chemical synthesis (11,12). However, this approach is limited by the availability of enzymes with the appropriate specificities.…”

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confidence: 99%

Peptide ligands for a sugar-binding protein isolated from a random peptide library.

Oldenburg

Loganathan

Goldstein

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

250

119

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Peptide Carbohydrate-protein interactions form the basis of a host of biological processes (1, 2). For example, the periplasmic monosaccharide-binding proteins of Gram-negative bacteria serve as receptors for transport and chemotaxis (3), while cell-surface lectins mediate such processes as cell-cell adhesion (4, 5) and lymphocyte migration through lymphoid tissues (6). Carbohydrate recognition is central to the enzymatic synthesis and degradation of polysaccharides, glycoproteins, and glycolipids that play essential roles in metabolism and in the maintenance of cellular structures. Carbohydrate-protein associations are also critical in certain cycles of viral infection. For example, binding of the hemagglutinin protein of human influenza virus to sialic acid residues on erythrocyte cell-surface glycoproteins represents the initial step in influenza infection (7).Consequently, the development of potent inhibitors of carbohydrate-specific proteins may be of considerable importance in the generation of new therapeutic agents. However, the synthesis of complex carbohydrate ligands and analogs often requires many time-consuming, low-yielding steps. Chemical synthesis of oligosaccharides requires sophisticated strategies for protecting/deprotecting and assembling sugar monomers and for controlling product regiochemistry and stereochemistry (8-10). Enzymatic synthesis using glycosyltransferases has emerged as a useful alternative to chemical synthesis (11,12). However, this approach is limited by the availability of enzymes with the appropriate specificities.An alternative approach to the synthesis of polysaccharide ligands for carbohydrate-specific receptors is to ask whether peptides can be found that bind these proteins with high affinities. We describe a strategy for identifying novel peptide ligands for carbohydrate-binding proteins based on the screening of a large, highly diverse peptide library expressed on the surface of filamentous phage fd (13-16). The library consists of phage bearing random octapeptides fused to the amino terminus of the minor coat protein, pIII, and was screened by affinity purification on immobilized receptor. We chose as a model system the lectin Con A fromjack bean, whose physicochemical properties have been extensively studied (17). Con A is a tetramer composed of four identical polypeptide chains consisting of 237 residues each. Con A, which interacts preferentially with oligosaccharides bearing terminal a-linked mannose or glucose residues, is frequently employed in the purification and structural characterization of carbohydrates and glycoproteins and is also a lymphocyte mitogen. We report the isolation of peptide ligands for Con A that prevent binding of known monosaccharide ligands of the lectin and that inhibit Con A-dependent precipitation of polysaccharides. The prospect that specific ligands for carbohydrate-binding proteins can be assembled by coupling amino acid building blocks (rather than sugars) should greatly reduce the synthetic difficulties associated with i...

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“…The advantages of using glycosyl transferases for oligosaccharide synthesis have been well reviewed [12,13] and arise mostly from the mild reaction conditions, the lack of requirement for protection and deprotection steps and the regio-and stereoselectivity of the glycosylation step.…”

Section: Introductionmentioning

confidence: 99%

Substrate and donor specificity of glycosyl transferases

Ernst¹,

Oehrlein²

1999

Glycotechnology

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It has been shown that all selectins recognize the carbohydrate epitopes sialyl Lewis x and sialyl Lewis a . For the establishment of the structure-activity relationship, the efficient synthesis of these tetrasaccharides and derivatives is therefore of vital interest. The glycosyl transferase-mediated approach is summarized with emphasis on the use of modified acceptors and modified sugar-nucleotide donors. A survey of the involved enzymes: b(1-3) and b(1-4)galactosyl transferases, a(2-3)sialyl transferase, FucT III and FucT VI reveals that the enzymatic synthesis is highly efficient for the rapid preparation of sialyl Lewis x -and sialyl Lewis a -derivatives.

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Enzyme-catalyzed synthesis of carbohydrates

Cited by 374 publications

References 217 publications

Chemical synthesis of GDP-fucose analogs and their utilization by the Lewis *A(1 → 4) fucosyltransferase

Chemical synthesis of GDP-fucose analogs and their utilization by the Lewis *A(1 → 4) fucosyltransferase

Peptide ligands for a sugar-binding protein isolated from a random peptide library.

Substrate and donor specificity of glycosyl transferases

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