Abstract
A glycomimetic oligonucleotide conjugate bearing four galactose residues on a mannose core is synthesized using oligonucleotide solid-phase synthesis and Cu(I)-catalyzed azide-alkyne 1,3-dipolar cycloaddition (CuAAC, or “click” chemistry). To achieve this purpose, new building blocks (including the solid support and phosphoramidites) are synthesized and used on a DNA synthesizer to generate a tetraalkyne oligonucleotide, which is then conjugated with a galactose azide derivative by click chemistry to afford the desired 3′-tetragalactosyl-mannose oligonucleotide conjugate. The procedures described in this chapter provide a general approach for the synthesis of novel glycoconjugates that can be immobilized to a DNA chip via DNA-directed immobilization to study, for example, their multivalent interactions with lectins in cellular targeting/uptake, etc.
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References
Varki, A. (1993) Biological Roles of Oligosaccharides – All of the Theories Are Correct. Glycobiology 3, 97–130.
Dwek, R. A. (1996) Glycobiology: Toward understanding the function of sugars. Chem. Rev. 96, 683–720.
Lis, H., and Sharon, N. (1998) Lectins: Carbohydrate-Specific Proteins That Mediate Cellular Recognition. Chem. Rev. 98, 637–674.
Imberty, A., Wimmerova, M., Mitchell, E. P., and Gilboa-Garber, N. (2004) Structures of the lectins from Pseudomonas aeruginosa: insights into the molecular basis for host glycan recognition. Microbes Infect. 6, 221–228.
Chemani, C., Imberty, A., de Bentzmann, S., Pierre, M., Wimmerova, M., Guery, B. P., and Faure, K. (2009) Role of LecA and LecB Lectins in Pseudomonas aeruginosa-Induced Lung Injury and Effect of Carbohydrate Ligands. Infect. Immun. 77, 2065–2075.
Deguise, I., Lagnoux, D., and Roy, R. (2007) Synthesis of glycodendrimers containing both fucoside and galactoside residues and their binding properties to Pa-IL and PA-IIL lectins from Pseudomonas aeruginosa. New J. Chem. 31, 1321–1331.
Lundquist, J. J., and Toone, E. J. (2002) The cluster glycoside effect. Chem. Rev. 102, 555–578.
Darbre, T., and Reymond, J. L. (2008) Glycopeptide dendrimers for biomedical applications. Curr. Top. Med. Chem. 8, 1286–1293.
Singh, Y., Renaudet, O., Defrancq, E., and Dumy, P. (2005) Preparation of a multitopic glycopeptide-oligonucleotide conjugate. Org. Lett. 7, 1359–1362.
Marra, A., Moni, L., Pazzi, D., Corallini, A., Bridi, D., and Dondoni, A. (2008) Synthesis of sialoclusters appended to calix[4]arene platforms via multiple azide-alkyne cycloaddition. New inhibitors of hemagglutination and cytopathic effect mediated by BK and influenza A viruses. Org. Biomol. Chem. 6, 1396–1409.
Cecioni, S., Lalor, R., Blanchard, B., Praly, J. P., Imberty, A., Matthews, S. E., and Vidal, S. (2009) Achieving high affinity towards a bacterial lectin through multivalent topological isomers of calix[4]arene glycoconjugates. Chem. Eur. J. 15, 13232–13240.
Dubber, M., and Lindhorst, T. K. (2001) Trehalose-based octopus glycosides for the synthesis of carbohydrate-centered PAMAM dedrimers and thiourea-bridged glycoclusters. Org. Lett. 3, 4019–4022.
Ortega-Munoz, M., Perez-Balderas, F., Morales-Sanfrutos, J., Hernandez-Mateo, F., Isac-Garcia, J., and Santoyo-Gonzalez, F. (2009) Click Multivalent Heterogeneous Neoglycoconjugates – Modular Synthesis and Evaluation of Their Binding Affinities. Eur. J. Org. Chem., 2454–2473.
Garcia-Lopez, J. J., Hernandez-Mateo, F., Isac-Garcia, J., Kim, J. M., Roy, R., Santoyo-Gonzalez, F., and Vargas-Berenguel, A. (1999) Synthesis of per-glycosylated beta-cyclodextrins having enhanced lectin binding affinity. J. Org. Chem. 64, 522–531.
Matsuura, K., Hibino, M., Yamada, Y., and Kobayashi, K. (2001) Construction of Glyco-Clusters by Self-Organization of Site-Specifically Glycosylated Oligonucleotides and Their Cooperative Amplification of Lectin-Recognition. J. Am. Chem. Soc. 123, 357–358.
Zatsepin, T. S., and Oretskaya, T. S. (2004) Synthesis and applications of oligonucleotide-carbohydrate conjugates. Chem. Biodiversity 1, 1401–1417.
Biessen, E. A. L., Vietsch, H., Rump, E. T., Fluiter, K., Kuiper, J., Bijsterbosch, M. K., and Van Berkel, T. J. C. (1999) Targeted delivery of oligodeoxynucleotides to parenchymal liver cells in vivo. Biochem. J. 340, 783–792.
Katajisto, J., Heinonen, P., and Lonnberg, H. (2004) Solid-phase synthesis of oligonucleotide glycoconjugates bearing three different glycosyl groups: Orthogonally protected bis(hydroxymethyl)-N,N′-bis(3-hydroxypropyl)malondiamide phosphoramidite as key building block. J. Org. Chem. 69, 7609–7615.
Katajisto, J., Virta, P., and Lonnberg, H. (2004) Solid-phase synthesis of multiantennary oligonucleotide glycoconjugates utilizing on-support oximation. Bioconjugate Chem. 15, 890–896.
Bouillon, C., Meyer, A., Vidal, S., Jochum, A., Chevolot, Y., Cloarec, J. P., Praly, J. P., Vasseur, J. J., and Morvan, F. (2006) Microwave assisted “click” chemistry for the synthesis of multiple labeled-carbohydrate oligonucleotides on solid support. J. Org. Chem. 71, 4700–4702.
Beaucage, S. L., and Caruthers, M. H. (1981) Deoxynucleoside phosphoramidites – a new class of key intermediates for deoxypolynucleotide synthesis. Tetrahedron Lett. 22, 1859–1862.
Tornoe, C. W., Christensen, C., and Meldal, M. (2002) Peptidotriazoles on solid phase: [1,2,3]-triazoles by regiospecific copper(I)-catalyzed 1,3-dipolar cycloadditions of terminal alkynes to azides. J. Org. Chem. 67, 3057–3064.
Rostovtsev, V. V., Green, L. G., Fokin, V. V., and Sharpless, K. B. (2002) A Stepwise Huisgen Cycloaddition Process: Copper(I)-Catalyzed Regioselective “Ligation” of Azides and Terminal Alkynes. Angew. Chem. Int. Ed. 41, 2596–2599.
Wacker, R., and Niemeyer, C. M. (2004) DDI-mu FIA – A readily configurable microarray-fluorescence immunoassay based on DNA-directed immobilization of proteins. Chembiochem 5, 453–459.
Chevolot, Y., Bouillon, C., Vidal, S., Morvan, F., Meyer, A., Cloarec, J. P., Jochum, A., Praly, J. P., Vasseur, J. J., and Souteyrand, E. (2007) DNA-based carbohydrate biochips: A platform for surface glyco-engineering. Angew. Chem. Int. Ed. 46, 2398–2402.
Pourceau, G., Meyer, A., Vasseur, J. J., and Morvan, F. (2009) Azide Solid Support for 3′-Conjugation of Oligonucleotides and Their Circularization by Click Chemistry. J. Org. Chem. 74, 6837–6842.
Hasegawa, T., Numata, M., Okumura, S., Kimura, T., Sakurai, K., and Shinkai, S. (2007) Carbohydrate-appended curdlans as a new family of glycoclusters with binding properties both for a polynucleotide and lectins. Org. Biomol. Chem. 5, 2404–2412.
Pourceau, G., Meyer, A., Vasseur, J. J., and Morvan, F. (2009) Synthesis of Mannose and Galactose Oligonucleotide Conjugates by Bi-click chemistry. J. Org. Chem. 74, 1218–1222.
Pourceau, G., Meyer, A., Vasseur, J. J., and Morvan, F. (2008) Combinatorial and automated synthesis of phosphodiester galactosyl cluster on solid support by click chemistry assisted by microwaves. J. Org. Chem. 73, 6014–6017.
Lonnberg, H. (2009) Solid-Phase Synthesis of Oligonucleotide Conjugates Useful for Delivery and Targeting of Potential Nucleic Acid Therapeutics. Bioconjugate Chem. 20, 1065–1094.
Pon, R. T., and Yu, S. Y. (1997) Hydroquinone-O,O′-diacetic acid (Q-linker) as a replacement for succinyl and oxalyl linker arms in solid phase oligonucleotide synthesis. Nucleic Acids Res. 25, 3629–3635.
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Pourceau, G., Meyer, A., Vasseur, JJ., Morvan, F. (2011). Synthesis of a Glycomimetic Oligonucleotide Conjugate by 1,3-Dipolar Cycloaddition. In: Mark, S. (eds) Bioconjugation Protocols. Methods in Molecular Biology, vol 751. Humana Press. https://doi.org/10.1007/978-1-61779-151-2_11
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DOI: https://doi.org/10.1007/978-1-61779-151-2_11
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