- Chemoenzymatic synthesis of 2-azidoethyl-ganglio-oligosaccharides GD3, GT3, GM2, GD2, GT2, GM1, and GD1a
-
We have synthesized several ganglio-oligosaccharide structures using glycosyltransferases from Campylobacter jejuni. The enzymes, α-(2→3/8)-sialyltransferase (Cst-II), β-(1→4)-N- acetylgalactosaminyltransferase (CgtA), and β-(1→3)- galactosyltransferase (CgtB), were produced in large-scale fermentation from Escherichia coli and further characterized based on their acceptor specificities. 2-Azidoethyl-glycosides corresponding to the oligosaccharides of GD3 (α-D-Neup5Ac-(2→8)-α-D-Neup5Ac-(2→3)-β-D-Galp- (1→4)-β-D-Glcp-), GT3 (α-D-Neup5Ac-(2→8)-α-D-Neup5Ac- (2→8)-α-D-Neup5Ac-(2→3)-β-D-Galp-(1→4)-β-D-Glcp-) , GM2 (β-D-GalpNAc-(1→4)-[α-D-Neup5Ac-(2→3)]-β-D-Galp- (1→4)-β-D-Glcp-), GD2 (β-D-GalpNAc-(1→4)-[α-D-Neup5Ac- (2→8)-α-D-Neup5Ac-(2→3)]-β-D-Galp-(1→4) -β-D-Glcp-), GT2 (β-D-GalpNAc-(1→4)-[α-D-Neup5Ac-(2→8) -α-D-Neup5Ac-(2→8)-α-D-Neup5Ac-(2→3)]-β-D-Galp- (1→4)-β-D-Glcp-), and GM1 (β-D-Galp-(1→3)-β-D-GalpNAc- (1→4)-[α-D-Neup5Ac-(2→3)]-β-D-Galp-(1→4) -β-D-Glcp-) were synthesized in high yields (gram-scale). In addition, a mammalian α-(2→3)-sialyltransferase (ST3Gal I) was used to sialylate GM1 and generate GD1a (α-D-Neup5Ac-(2→3)-β-D-Galp-(1→3)- β-D-GalpNAc-(1→4)-[α-D-Neup5Ac-(2→3)]-β-D-Galp- (1→4)-β-D-Glcp-) oligosaccharide. We also cloned and expressed a rat UDP-N-acetylglucosamine-4′epimerase (GalNAcE) in E. coli AD202 cells for cost saving in situ conversion of less expensive UDP-GlcNAc to UDP-GalNAc.
- Blixt, Ola,Vasiliu, Daniela,Allin, Kirk,Jacobsen, Nathan,Warnock, Dawn,Razi, Nahid,Paulson, James C.,Bernatchez, Stephane,Gilbert, Michel,Wakarchuk, Warren
-
-
Read Online
- Design and synthesis of cluster neoglycoconjugates based on D-glucose
-
Synthetic approaches have been developed to tetravalent neoglycoconjugates with different structures of the hydrophobic fragment and D-glucose fragment as branching core. The syntheses have been accomplished by preparation of blocks with terminal C C triple bonds and hydrophobic fragment and hydrophilic component (lactose derivative), followed by their conjugation.
- Kurochkina,Budanova,Sebyakin, Yu. L.
-
-
Read Online
- The trans-sialidase from Trypanosoma cruzi efficiently transfers α-(2→3)-linked N-glycolylneuraminic acid to terminal β-galactosyl units
-
The trans-sialidase from Trypanosoma cruzi (TcTS), the agent of Chagas' disease, is a unique enzyme involved in mammalian host-cell invasion. Since T. cruzi is unable to synthesize sialic acids de novo, TcTS catalyzes the transfer of α-(2→3)-sialyl residues from the glycoconjugates of the host to terminal β-galactopyranosyl units present on the surface of the parasite. TcTS also plays a key role in the immunomodulation of the infected host. Chronic Chagas' disease patients elicit TcTS-neutralizing antibodies that are able to inhibit the enzyme. N-Glycolylneuraminic acid has been detected in T. cruzi, and the trans-sialidase was pointed out as the enzyme involved in its incorporation from host glycoconjugates. However, N-glycolylneuraminic acid α-(2→3)-linked-containing oligosaccharides have not been analyzed as donors in the T. cruzi trans-sialidase reaction. In this paper we studied the ability of TcTS to transfer N-glycolylneuraminic acid from Neu5Gc(α2→3)Gal(β1→4)GlcβOCH2CH2N3 (1) and Neu5Gc(α2→3)Gal(β1→3)GlcNAcβOCH2CH2N3 (2) to lactitol, N-acetyllactosamine and lactose as acceptor substrates. Transfer from 1 was more efficient (50-65%) than from 2 (20-30%) for the three acceptors. The reactions were inhibited when the enzyme was preincubated with a neutralizing antibody. Km values were calculated for 1 and 2 and compared with 3′-sialyllactose using lactitol as acceptor substrate. Analysis was performed by high-performance anion-exchange (HPAEC) chromatography. A competitive transfer reaction of compound 1 in the presence of 3′-sialyllactose and N-acetyllactosamine showed a better transfer of Neu5Gc than of Neu5Ac.
- Agusti, Rosalia,Giorgi, Maria Eugenia,de Lederkremer, Rosa M.
-
-
Read Online
- Synthesis of DNP-modified GM3-based anticancer vaccine and evaluation of its immunological activities for cancer immunotherapy
-
Tumor-associated carbohydrate antigens (TACAs) are attractive targets for vaccine development. In this context, we described a strategy combining artificial TACA and glycoengineering for cancer vaccine development. A 2,4-ditrophenyl (DNP)-modified GM3 int
- Lin, Han,Hong, Haofei,Feng, Lipeng,Shi, Jie,Zhou, Zhifang,Wu, Zhimeng
-
supporting information
p. 4041 - 4044
(2021/06/15)
-
- The design and synthesis of an α-Gal trisaccharide epitope that provides a highly specific anti-Gal immune response
-
Carbohydrate antigens displaying Galα(1,3)Gal epitopes are recognized by naturally occurring antibodies in humans. These anti-Gal antibodies comprise up to 1% of serum IgG and have been viewed as detrimental as they are responsible for hyperacute organ rejections. In order to model this condition, α(1,3)galactosyltransferase-knockout mice are inoculated against the Galα(1,3)Gal epitope. In our study, two α-Gal trisaccharide epitopes composed of either Galα(1,3)Galβ(1,4)GlcNAc or Galα(1,3)Galβ(1,4)Glc linked to a squaric acid ester moiety were examined for their ability to elicit immune responses in KO mice. Both target epitopes were synthesized using a two-component enzymatic system using modified disaccharide substrates containing a linker moiety for coupling. While both glycoconjugate vaccines induced the required high anti-Gal IgG antibody titers, it was found that this response had exquisite specificity for the Galα(1,3)Galβ(1,4)GlcNAc hapten used, with little cross reactivity with the Galα(1,3)Galβ(1,4)Glc hapten. Our findings indicate that while homogenous glycoconjugate vaccines provide high IgG titers, the carrier and adjuvanting factors can deviate the specificity to an antigenic determinant outside the purview of interest.
- Anraku, Kensaku,Sato, Shun,Jacob, Nicholas T.,Eubanks, Lisa M.,Ellis, Beverly A.,Janda, Kim D.
-
p. 2979 - 2992
(2017/04/10)
-
- Synthesis and antibacterial activities of novel tyrocidine A glycosylated derivatives towards multidrug-resistant pathogens
-
Glycosylation can have a multifaceted impact on the properties and functions of peptides and plays a critical role in interacting with or binding to the target molecules. Herein, based on the previously reported method for macrocyclic glycopeptide synthesis, two series of tyrocidine A glycosylated derivatives (1a-f and 2a-f) were synthesized and evaluated for their antibacterial activities to further study the structure and activity relationships (SAR). Biological studies showed that the synthetic glycosylated derivatives had good antibacterial activities towards methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus. SAR studies based on various glycans and linkages were used to enhance the biochemical profile, resulting in the identification of several potent antibiotics, such as 1f, with a great improved therapeutic index than tyrocidine A.
- Zou, Yan,Zhao, Qingjie,Zhang, Chunmei,Wang, Liang,Li, Wenjuan,Li, Xiang,Wu, Qiuye,Hu, Honggang
-
p. 586 - 592
(2015/07/02)
-
- MODULAR SYNTHESIS OF AMPHIPHILIC JANUS GLYCODENDRIMERS AND THEIR SELF-ASSEMBLY INTO GLYCODENDRIMERSOMES
-
The invention concerns compounds of the formula (I) wherein: Y1 and Y2 are independently a monosaccharide or disaccharide; X1 and X2 are independently -(R9-O)m-, -(R10)P-, -O-(R11-O)q-, -R16-O-R17-O- or a covalent bond; Q1 and Q2 are independently a nitrogen-containing heterocycle moiety; Z1 and Z2 are independently -(O-R7)-, -(O-C(=O)-R8)a-, -O-C(=O)-R12-C(=0)-R13-, -O- C(=O)-R14-C(=O)-R15 or a covalent bond; R7-R17 are each independently C1-C6 alkyl; R1-R6 are each independently a linear or branched alkly group; b, c, d, e, f, and g are 0 or 1, provided b + c + d equals at least 2 and e + f + g equals at least 2; and a, m, p, and q are each an integer from 1-6.
- -
-
Paragraph 00132
(2014/12/12)
-
- Modular synthesis of amphiphilic Janus glycodendrimers and their self-assembly into glycodendrimersomes and other complex architectures with bioactivity to biomedically relevant lectins
-
The modular synthesis of 7 libraries containing 51 self-assembling amphiphilic Janus dendrimers with the monosaccharides d-mannose and d-galactose and the disaccharide d-lactose in their hydrophilic part is reported. These unprecedented sugar-containing dendrimers are named amphiphilic Janus glycodendrimers. Their self-assembly by simple injection of THF or ethanol solution into water or buffer and by hydration was analyzed by a combination of methods including dynamic light scattering, confocal microscopy, cryogenic transmission electron microscopy, Fourier transform analysis, and micropipet-aspiration experiments to assess mechanical properties. These libraries revealed a diversity of hard and soft assemblies, including unilamellar spherical, polygonal, and tubular vesicles denoted glycodendrimersomes, aggregates of Janus glycodendrimers and rodlike micelles named glycodendrimer aggregates and glycodendrimermicelles, cubosomes denoted glycodendrimercubosomes, and solid lamellae. These assemblies are stable over time in water and in buffer, exhibit narrow molecular-weight distribution, and display dimensions that are programmable by the concentration of the solution from which they are injected. This study elaborated the molecular principles leading to single-type soft glycodendrimersomes assembled from amphiphilic Janus glycodendrimers. The multivalency of glycodendrimersomes with different sizes and their ligand bioactivity were demonstrated by selective agglutination with a diversity of sugar-binding protein receptors such as the plant lectins concanavalin A and the highly toxic mistletoe Viscum album L. agglutinin, the bacterial lectin PA-IL from Pseudomonas aeruginosa, and, of special biomedical relevance, human adhesion/growth-regulatory galectin-3 and galectin-4. These results demonstrated the candidacy of glycodendrimersomes as new mimics of biological membranes with programmable glycan ligand presentations, as supramolecular lectin blockers, vaccines, and targeted delivery devices.
- Percec, Virgil,Leowanawat, Pawaret,Sun, Hao-Jan,Kulikov, Oleg,Nusbaum, Christopher D.,Tran, Tam M.,Bertin, Annabelle,Wilson, Daniela A.,Peterca, Mihai,Zhang, Shaodong,Kamat, Neha P.,Vargo, Kevin,Moock, Diana,Johnston, Eric D.,Hammer, Daniel A.,Pochan, Darrin J.,Chen, Yingchao,Chabre, Yoann M.,Shiao, Tze C.,Bergeron-Brlek, Milan,Andre, Sabine,Roy, Rene,Gabius, Hans-J.,Heiney, Paul A.
-
supporting information
p. 9055 - 9077
(2013/07/26)
-
- Site-selective modification of proteins for the synthesis of structurally defined multivalent scaffolds
-
A combination of classical site-directed mutagenesis, genetic code engineering and bioorthogonal reactions delivered a chemically modified barstar protein with one or four carbohydrates installed at specific residues. These protein conjugates were employed in multivalent binding studies, which support the use of proteins as structurally defined scaffolds for the presentation of multivalent ligands.
- Artner, Lukas M.,Merkel, Lars,Bohlke, Nina,Beceren-Braun, Figen,Weise, Christoph,Dernedde, Jens,Budisa, Nediljko,Hackenberger, Christian P. R.
-
supporting information; experimental part
p. 522 - 524
(2012/02/06)
-
- Enzymatic synthesis of a 6-sialyl lactose analogue using a pH-responsive water-soluble polymer support
-
The Letter describes a strategy for the enzymatic synthesis of glycans based on a pH-responsive water-soluble polymer. In neutral condition, the polymer is water-soluble and convenient for in-solution enzymatic synthesis, whereas in acidic condition (pH lower than 4.0), the polymer disconnects with the product and becomes insoluble, which can be easily removed. A 6-Sialyl lactose analogue was synthesized as a model reaction using this approach.
- Wang, Wenjun,Li, Lei,Jin, Chen,Niu, Yujie,Li, Sen,Ma, Ji,Li, Linfeng,Liu, Yu,Cai, Li,Zhao, Wei,Wang, Peng George
-
supporting information; experimental part
p. 5041 - 5044
(2011/10/09)
-
- Preparation of oligosaccharides by homogenous enzymatic synthesis and solid phase extraction
-
This communication describes a method for enzymatic preparation of bioactive glycans, which integrated the high-efficiency of homogenous phase enzymatic reaction and fast separation of solid phase extraction.
- Wang, Wenjun,Jin, Chen,Guo, Lina,Liu, Yu,Wan, Yue,Wang, Xin,Li, Lei,Zhao, Wei,Wang, Peng George
-
supporting information; experimental part
p. 11240 - 11242
(2011/12/05)
-
- Preparation of aminoethyl glycosides for glycoconjugation
-
The synthesis of a number of aminoethyl glycosides of cell-surface carbohydrates, which are important intermediates for glycoarray synthesis, is described. A set of protocols was developed which provide these intermediates, in a short number of steps, from commercially available starting materials.
- Sardzik, Robert,Noble, Gavin T.,Weissenborn, Martin J.,Martin, Andrew,Webb, Simon J.,Flitsch, Sabine L.
-
supporting information; experimental part
p. 699 - 703
(2011/01/03)
-
- Target-specific chemical acylation of lectins by ligand-tethered DMAP catalysts
-
Because sugar-binding proteins, so-called lectins, play important roles in many biological phenomena, the lectin-selective labeling should be useful for investigating biological processes involving lectins as well as providing molecular tools for analysis of saccharides and these derivatives. We describe herein a new strategy for lectin-selective labeling based on an acyl transfer reaction directed by ligand-tethered DMAP (4-dimethylaminopyridine). DMAP is an effective acyl transfer catalyst, which can activate an acyl ester for its transfer to a nucleophilic residue. To direct the acyl transfer reaction to a lectin of interest, we attached the DMAP to a saccharide ligand specific for the target lectin. It was clearly demonstrated by biochemical analyses that the target-selective labeling of Congerin II, an animal lectin having selective affinity for Lactose/LacNAc (N-acetyllactosamine), was achieved in the presence of Lactethered DMAPs and acyl donors containing probes such as fluorescent molecules or biotin. Conventional peptide mapping experiments using HPLC and tandem mass-mass analysis revealed that the acyl transfer reaction site-specifically occurred at Tyr 51 of Cong II. This strategy was successfully extended to other lectins by changing the ligand part of the ligand-tethered DMAP. We also demonstrated that this labeling method is applicable not only to purified lectin in test tubes, but also to crude mixtures such as E. coli lysates or homogenized animal tissue samples expressing Congerin.
- Koshi, Yoichiro,Nakata, Eiji,Miyagawa, Masayoshi,Tsukiji, Shinya,Ogawa, Tomohisa,Hamachi, Itaru
-
p. 245 - 251
(2008/10/09)
-
- Carbohydrate microarrays for assaying galactosyltransferase activity
-
Carbohydrate microarrays have been used recently for the rapid analysis of glycan-protein or glycan-cell interactions and for the detection of pathogens. As a demonstration of its significance and versatility, the microarray technology has been applied in this effort to assay glycosyltransferase activities. In addition, carbohydrate microarray based methods have been employed to quantitatively determine binding affinities between lectins and carbohydrates.
- Park, Sungjin,Shin, Injae
-
p. 1675 - 1678
(2008/02/02)
-
- Preparation of carbohydrate-oligonucleotide conjugates using the squarate spacer
-
Attachment of carbohydrates to oligonucleotides has proven to induce receptor-mediated endocytosis. A facile method for the formation of covalent linkages between glycans and oligonucleotides is herein described. Thus, use of 3,4-diethoxy-3-cyclobutene-1,2-dione as a linking reagent provides easy conjugation between carbohydrates bearing an amino group at the reducing end and oligonucleotides bearing an aminoalkyl modification.
- Yan, Hongbin,Aguilar, Aime Lopez,Zhao, Yuyan
-
p. 6535 - 6538
(2008/03/18)
-
- An efficient glycosylation reaction for the synthesis of asialo GM2 analogues
-
We investigated the coupling reaction of glycosyl donors N-trichloroethoxycarbonyl-galactosamine-O-trichloroacetimidate (2a) and N-p-nitrobenzyloxycarbonyl-galactosamine-O-trichloroacetimidate (2b) with the 4′-OH of lactose derivatives (3a-d) to synthesiz
- Sun, Bin,Pukin, Aliaksei V.,Visser, Gerben M.,Zuilhof, Han
-
p. 7371 - 7374
(2007/10/03)
-
- Synthesis and enzyme-specific activation of carbohydrate-geldanamycin conjugates with potent anticancer activity
-
Geldanamycin (GA) is a potent anticancer antibiotic that inhibits Hsp90. Its potential clinical utility is hampered by its severe toxicity. To alleviate this problem, we synthesized a series of carbohydrate-geldanamycin conjugates for enzyme-specific activation to increase tumor selectivity. The conjugation was carried out at the C-17-position of GA. Their anticancer activity was tested in a number of cancer cell lines. The enzyme-specific activation of these conjugates was evaluated with β-galactosidase and β-glucosidase. Evidently, glycosylation of C-17-position converted GA to an inactive prodrug before enzyme cleavage. Glucose-GA, as positive control, showed anticancer activity with IC50 of 70.2-380.9 nM in various cancer cells by β-glucosidase activation inside of the tumor cells, which was confirmed by 3-fold inhibition using β-glucosidase specific inhibitor [2,5-dihydroxyniethy-3,4-dihydroxypyrrolidine (DMDP)]. Compared to glucose-GA, galactose- and lactose-GA conjugates exhibited much less activity with IC 50 greater than 8000-25 000 nM. However, when galactose- and lactose-GA were incubated with β-galactosidase in the cells, their anticancer activity was enhanced by 3- to 40-fold. The results suggest that GA can be inactivated by glycosylation of C-17-position and reactivated for anticancer activity by β-galactosidase. Therefore, galactose-GA can be exploited in antibody-directed enzyme prodrug therapy (ADEPT) with β-galactosidase for enzyme-specific activation in tumors to increase tumor selectivity.
- Cheng, Hao,Cao, Xianhua,Xian, Ming,Fang, Lanyan,Cai, Tingwei Bill,Ji, Jacqueline Jia,Tunac, Josefino B.,Sun, Duxin,Wang, Peng George
-
p. 645 - 652
(2007/10/03)
-
- Synthesis and molecular recognition of carbohydrate-centered multivalent glycoclusters by a plant lectin RCA120
-
Water soluble and lectin-recognizable carbohydrate-centered glycoclusters were prepared efficiently by the Huisgen 1,3-cycloaddition reaction of methyl-2,3,4,6-tetra-O-propargyl β-d-galactopyranoside with 2-azidoethyl glycosides of lactose and N-acetyllac
- Gao, Yongjun,Eguchi, Atsuko,Kakehi, Kazuaki,Lee, Yuan C.
-
p. 6151 - 6157
(2007/10/03)
-
- Synthesis of aminoethyl glycosides of the ganglioside GM1 and asialo-GM1 oligosaccharide chains
-
4′-O-Glycosylation of 2-azidoethyl 2,3,6-tri-O-benzyl-4-O-(2,3-di-O- benzyl-6-O-benzoyl-β-D-galactopyranosyl)-β-D-glucopyranoside with a disaccharide donor, 4-trichloroacetamidophenyl 4,6-di-O-acetyl-2-deoxy-3-O-(2,3, 4,6-tetra-O-acetyl-β-D-galactopyranos
- Cheshev,Khatuntseva,Tsvetkov,Shashkov,Nifantiev
-
-
- Design and synthesis of biotin chain-terminated glycopolymers for surface glycoengineering
-
Biotin chain-terminated glycopolymers were generated by cyanoxyl-mediated free-radical polymerization using a biotin-derivatized arylamine initiator with high conversion (75%) and low polydispersity (1.30). Streptavidin-biotinylated glycopolymer binding w
- Sun, Xue-Long,Faucher, Keith M.,Houston, Michelle,Grande, Daniel,Chaikof, Elliot L.
-
p. 7258 - 7259
(2007/10/03)
-
- Phage-display selection of a human single-chain Fv antibody highly specific for melanoma and breast cancer cells using a chemoenzymatically synthesized GM3-carbohydrate antigen
-
Overexpression of the cell-surface glycosphingolipid GM3 is associated with a number of different cancers, including those of the skin, colon, breast, and lung. Antibodies against the GM3 epitope have potential application as therape
- Lee, Kyung Joo,Mao, Shenlan,Sun, Chengzao,Gao, Changshou,Blixt, Ola,Arrues, Sandra,Hom, Louis G.,Kaufmann, Gunnar F.,Hoffman, Timothy Z.,Coyle, Avery R.,Paulson, James,Felding-Habermann, Brunhilde,Janda, Kim D.
-
p. 12439 - 12446
(2007/10/03)
-
- Synthesis of the Lewis b hexasaccharide and squarate acid-HSA conjugates thereof with various saccharide loadings
-
The Lewis b hexasaccharide, α-Ll-Fucp-(1 → 2)-β-D-Galp-(1 → 3)-[α-L-Fucp-(1 → 4)]-β-D-GlcpNAc-(1 → 3)-β-D-Galp-(1 → 4)-β-D-Glcp, has been synthesised using a convergent synthesis. Starting from ethyl 4,6-O-benzylidene-2-deoxy-2-phthalimido-1-thio-β-D-gluc
- Chernyak, Anatoly,Oscarson, Stefan,Turek, Dominika
-
p. 309 - 316
(2007/10/03)
-