Synthesis of Solid-Supported Mirror-Image Sugars: A Novel Method for Selecting Receptors for Cellular-Surface Carbohydrates

Kozlov, I. A.; Mao, Shenlan; Huang, Xuefei; Lee, Lac; Sears, P.S.; Gao, Changshou; Coyle, Avery R.; Janda, Kim D.; Wong, Chi Huey

doi:10.1002/1439-7633(20011001)2:10<741::aid-cbic741>3.0.co;2-b

Cited by 33 publications

(23 citation statements)

References 26 publications

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“…To investigate the underlying molecular basis, we determined the crystal structures of D-sialic acid aldolase and RS-aldolase. All mutations are away from the catalytic center, except for V251I, which is near the opening of the (␣/␤) 8 -barrel and proximal to the Schiff base-forming Lys-165. The change of specificity from D-sialic acid to RS-aldolase can be attributed mainly to the V251I substitution, which creates a narrower sugar-binding pocket, but without altering the chirality in the reaction center.…”

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Modulation of Substrate Specificities of d-Sialic Acid Aldolase through Single Mutations of Val-251

Chou

et al. 2011

Journal of Biological Chemistry

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In a recent directed-evolution study, Escherichia coli D-sialic acid aldolase was converted by introducing eight point mutations into a new enzyme with relaxed specificity, denoted RSaldolase (also known formerly as L-3-deoxy-manno-2-octulosonic acid (L-KDO) aldolase), which showed a preferred selectivity toward L-KDO. To investigate the underlying molecular basis, we determined the crystal structures of D-sialic acid aldolase and RS-aldolase. All mutations are away from the catalytic center, except for V251I, which is near the opening of the (␣/␤) 8 -barrel and proximal to the Schiff base-forming Lys-165. The change of specificity from D-sialic acid to RS-aldolase can be attributed mainly to the V251I substitution, which creates a narrower sugar-binding pocket, but without altering the chirality in the reaction center. The crystal structures of D-sialic acid aldolase⅐L-arabinose and RS-aldolase⅐hydroxypyruvate complexes and five mutants (V251I, V251L, V251R, V251W, and V251I/V265I) of the D-sialic acid aldolase were also determined, revealing the location of substrate molecules and how the contour of the active site pocket was shaped. Interestingly, by mutating Val251 alone, the enzyme can accept substrates of varying size in the aldolase reactions and still retain stereoselectivity. The engineered D-sialic acid aldolase may find applications in synthesizing unnatural sugars of C 6 to C 10 for the design of antagonists and inhibitors of glycoenzymes.Carbohydrates on cell surfaces play a pivotal role in the molecular recognition processes in various cellular interactions. Interfering with the recognition processes between pathogens and hosts may present therapeutic strategies for infectious diseases (1, 2). D-Sialic acids are widely found in nature, from bacteria to plants and animal tissues. Cell-surface glycoproteins and glycolipids frequently contain D-sialic acids at the termini of the oligosaccharide chains. The invading bacteria and viruses also target the cell-surface sialic acids of their hosts, which constitutes an important step for infectivity (3). On the other hand, the monosaccharide D-3-deoxy-manno-2-octulosonic acid (D-KDO) 2 has never been identified in vertebrates, but it is an important component in Gram-negative bacteria (4 -6). Because D-KDO is vital to the structural stability of bacterial outer membrane, targeting D-KDO and the related biosynthetic enzymes may be a new strategy for future antibacterial agent discovery.Currently, unnatural enantiomeric compounds become increasingly important in pharmaceutical implementations. Enantiomers (D-form) of natural peptides have been used to target cell-surface D-sugars. These D-peptide cell-surface sugar binders might serve as new drug candidates for infectious disease (7) and as tools for studying the interactions between proteins and carbohydrate. Owing to the resistance to hydrolases and high binding affinity to sugars, the unnatural D-peptides can become potential therapeutic agents. Through mirror-image phage display, natural L-peptides can be...

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Modulation of Substrate Specificities of d-Sialic Acid Aldolase through Single Mutations of Val-251

Chou

et al. 2011

Journal of Biological Chemistry

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“…These enantiomeric carbohydrates have been used as affinity ligands for the identification of high-affinity peptides that recognize L-KDO and L-sialic acid as an effort to develop a general strategy for use to develop Dpeptides to target cell-surface carbohydrates [4].…”

Section: D-peptides That Target Bacterial Cell-surface Carbohydratesmentioning

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Synthesis of Carbohydrate‐Based Antibiotics

et al. 2003

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Tackling the problem of carbohydrate recognition is a subject of current interest. Our strategies in this regard include development of new methods for the synthesis of saccharides and sugar arrays for screening, design of D-peptides to target bacterial cell-surface sugars, preparation of mechanism-based inhibitors of the enzymes involved in the biosynthesis of saccharides and the synthesis of aminoglycoside mimetics to target RNA.

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“…Oxidation of 4 with NaIO 4 gave a dial derivative (5). Finally, compound 5 was hydrolyzed to separate arabinose (6). The absolute configuration of 6 was verified as being the Dform using the Hara method.…”

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“…The absolute configuration of 6 was verified as being the Dform using the Hara method. On the other hand, L-NeuAc (7), which was synthesized from L-glucose using the authentic method, 6) gave L-arabinose in the same manner as 1. Thus the fact that D(L)-sialic acid could be converted to D(L)-arabinose was confirmed, as shown in Fig.…”

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Determination of the Absolute Configuration of Sialic Acids in Gangliosides from the Sea Cucumber Cucumaria echinata

Kisa

Yamada

Miyamoto

et al. 2007

CHEMICAL & PHARMACEUTICAL BULLETIN

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Gangliosides, an important group of glycosphingolipids due to their biological functions, 1) are characterized by the presence of one or more moles of sialic acid, N-acetylneuraminic acid (NeuAc) or N-glycolylneuraminic acid (NeuGc), in their carbohydrate parts. In general, naturally occurring sialic acids have been believed to be the D-form on the basis of biosynthetic evidence, i.e., NeuAc is synthesized from pyruvic acid and N-acetyl-D-mannosamine enzymatically. 2)However, there has been no report on the determination of the absolute configuration of the sialic acid residues in gangliosides. We attempted to determine the absolute configuration (D or L) of the sialic acid residues in the gangliosides 3,4) from the sea cucumber Cucumaria echinata.For the determination of the absolute configuration of aldoses in gangliosides, we have used the Hara method.5) However, this method cannot be applied to sialic acids directly. Therefore we prepared aldose from sialic acid, preserving the absolute configuration of the parent sialic acid, and then applied the Hara method to the aldose. As shown in Fig. 1, D(L)-arabinose should be obtained from D(L)-sialic acid in the process. First, this assumption was proved.D-NeuAc (1) was acetylated to give pentaacetate (2), which was methanolyzed with HCl-MeOH to yield methyl 2-O-methylneuraminate (3). Protection of the diol in the side chain of 3 with acetone afforded an isopropyridene derivative (4). Oxidation of 4 with NaIO 4 gave a dial derivative (5). Finally, compound 5 was hydrolyzed to separate arabinose (6). The absolute configuration of 6 was verified as being the Dform using the Hara method. On the other hand, L-NeuAc (7), which was synthesized from L-glucose using the authentic method, 6) gave L-arabinose in the same manner as 1. Thus the fact that D(L)-sialic acid could be converted to D(L)-arabinose was confirmed, as shown in Fig. 1.Next, this method was applied to the gangliosides SJG-1, CG-1, CEG-3, CEG-4, CEG-5, CEG-6, HLG-3, CEG-8, and CEG-9 3,4) from the sea cucumber C. echinata. The nine gangliosides were each methanolyzed and the sialic acid derivatives (3) produced were converted to ketals (4). Periodic acid oxidation of 4 followed by acidic hydrolysis of the oxidation products 5 gave arabinoses (6). Compound 6, obtained from each ganglioside, was determined to be the D-form using the Hara method, as shown in Fig. 2. Accordingly, the sialic acid residues (NeuAc and NeuGc) in the gangliosides from C. echinata must be the D-form. To the best of our knowledge, this is the first report of the determination of the absolute configuration of the sialic acid Determination of the Absolute Configuration of Sialic Acids inGangliosides from the Sea Cucumber Cucumaria echinata Fumiaki KISA, Koji YAMADA, Tomofumi MIYAMOTO, Masanori INAGAKI, and Ryuichi HIGUCHI* Faculty of Pharmaceutical Sciences, Kyushu University; 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. Received March 15, 2007; accepted April 6, 2007 Enantiomeric pairs of sialic acid, D-and L-NeuAc (N-...

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Synthesis of Solid-Supported Mirror-Image Sugars: A Novel Method for Selecting Receptors for Cellular-Surface Carbohydrates

Cited by 33 publications

References 26 publications

Modulation of Substrate Specificities of d-Sialic Acid Aldolase through Single Mutations of Val-251

Modulation of Substrate Specificities of d-Sialic Acid Aldolase through Single Mutations of Val-251

Synthesis of Carbohydrate‐Based Antibiotics

Determination of the Absolute Configuration of Sialic Acids in Gangliosides from the Sea Cucumber Cucumaria echinata

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