Glycobiology
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  • Glycobiology

    New England Biolabs (NEB) offers a selection of endoglycosidases, exoglycosidases, and heparinases for glycobiology research. Many of these reagents are recombinant, and all undergo several quality control assays, enabling us to provide products with lower unit cost, high purity and reduced lot-to-lot variation. All of our glycosidases are tested for contaminants. Since p-nitrophenyl-glycosides are not hydrolyzed by some exoglycosidases, we use only fluorescently-labeled oligosaccharides to screen for contaminating glycosidases.

    • Overview of Glycobiology

      Learn about the core sequences and common modifications of N-linked and O-linked glycans in this video. Analysis of these glycans and/or peptide portions of the glycoprotein can be accomplished with the use of deglycosylation enzymes, which are explained in detail. Unlike other chemical deglycosylation methods, enzymatic treatment is much gentler and can provide complete sugar removal with no protein degradation.

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    • Identification and Characterization of Protein Glycosylation

      Here we illustrate the use of glycosidases for the analysis of a model glycoprotein: recombinant human chorionic gonadotropin beta (hCGβ), which carries both N-glycans and O-glycans in this video. The technique requires only simple instrumentation and typical consumables, and it can be readily adapted to the analysis of multiple glycoprotein samples.

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    • Elucidating the Complexity of Heparin Oligosaccharide Analysis

      Learn more about the structural elucidation of complex and diverse heparin oligosaccharides using Bacteroides Heparinase I, II and III in combination with downstream mass spec analysis.

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    Glycobiology includes these subcategories:

    Endoglycosidases
    Exoglycosidases
    Heparinases
    Heparin Hexasaccharide Mass Spectrometry Standards

      Publications related to Glycobiology:

    1. Wong-Madden, S.T., Landry, D., and Guthrie, E.P. (1997) Discovery and Uses of Novel Glycosidases Techniques in Glycobiology 401-408.
    2. Kalidas, C., Joshi, L., Batt, C. (2001) Characterization of glycosylated variants of beta-lactoglobulin expressed in Pichia pastoris Protein Eng. 14, 201-7. PubMedID: 11342718
    3. Whistler, J.L., Enquist, J., Marley, A., Fong, J., Gladher, F., Tsuruda, P., Murray, S.R., Von Zastrow, M. (2002) Modulation of postendocytic sorting of G protein-coupled receptors Science 297, 615-20. PubMedID: 12142540
    4. Lüftenegger, D., Picard-Maureau, M., Stanke, N., Rethwilm, A., Lindemann, D. (2005) Analysis and function of prototype foamy virus envelope N glycosylation J. Virol.  79, 7664-72. PubMedID: 15919919
    5. Cipollo, J.F., Awad, A.M., Costello, C.E., Hirschberg, C.B. (2005) N-Glycans of Caenorhabditis elegans are specific to developmental stages J. Biol. Chem. 280, 26063-72. PubMedID: 15899899
    6. Srikrishna, G., Brive, L., Freeze, H.H. (2005) Novel carboxylated N-glycans contain oligosaccharide-linked glutamic acid Biochem. Biophys. Res. Commun. 332, 1020-7. PubMedID: 15922305
    7. Hagen, B.M., Sanders, K.M. (2006) Deglycosylation of the beta1-subunit of the BK channel changes its biophysical properties Am. J. Physiol.Cell. Physiol.  1, 750-6. PubMedID: 16738006
    8. O’Reilly, M.K., Zhang, G., Imperiali, B. (2006) In vitro evidence for the dual function of Alg2 and Alg11: essential mannosyltransferases in N-linked glycoprotein biosynthesis Biochemistry 8, 9593-603. PubMedID: 16878994
    9. Sielaff I. et al. (2006) Protein function microarrays based on self-immobilizing and self-labeling fusion proteins ChemBioChem. 7, 194-202. PubMedID: 16342318
    10. Eichler, R., Lenz, O., Garten, W., Strecker, T. (2006) The role of single N-glycans in proteolytic processing and cell surface transport of the Lassa virus glycoprotein.GP-C. Virol. J. 3, 41. PubMedID: 16737539
    11. Aoki, K., Perlman, M., Lim, J.M., Cantu, R., Wells, L., Tiemeyer, M. (2007) Dynamic developmental elaboration of N-linked glycan complexity in the Drosophila melanogaster embryo J. Biol. Chem. 282, 9127-42. PubMedID: 17264077
    12. Song, Y., Choi, M.H., Park, J.N., Kim, M.W., Kim, E.J., Kang, H.A., Kim, J.Y. (2007) Engineering of the yeast Yarrowia lipolytica for the production of glycoproteins lacking the outer-chain mannose residues of N-glycans Appl. Environ. Microbiol.  73, 4446-54. PubMedID: 17513593
    13. Teuton, J.R., Brandt, C.R. (2007) Sialic acid on herpes simplex virus type 1 envelope glycoproteins is required for efficient infection of cells  J. Virol. 81, 3731-9. PubMedID: 17229687
    14. Macnaughtan, M.A., Tian, F., Liu, S., Meng, L., Park, S., Azadi, P., Moremen, K.W., Prestegard, J.H. (2008) 13C-sialic acid labeling of glycans on glycoproteins using ST6Gal-I J. Am. Chem Soc. 130, 11864-5. PubMedID: 18700760
    15. Banala J. et al. (2008) Caged substrates for protein labeling and immobilization Chembiochem 4, 38-41. PubMedID: 18033718
    16. Baudino L, Shinohara Y, Nimmerjahn F, Furukawa J, Nakata M, Martínez-Soria E, Petry F, Ravetch JV, Nishimura S, Izui S. (2008) Crucial role of aspartic acid at position 265 in the CH2 domain for murine IgG2a and IgG2b Fc-associated effector functions J Immunol. 181(9), 6664-9. PubMedID: 18941257
    17. Oh, D.B., Park, J.S., Kim, M.W., Cheon, S.A., Kim, E.J., Moon, H.Y., Kwon, O., Rhee, S.K., Kang, H.A. (2008) Glycoengineering of the methylotrophic yeast Hansenula polymorpha for the production of glycoproteins with trimannosyl core N-glycan by blocking core oligosaccharide assembly Biotechnol. J. 3, 659-68. PubMedID: 18320567
    18. Zhuo, Y., Chammas, R., Bellis, S.L. (2008) Sialylation of beta1 integrins blocks cell adhesion to galectin-3 and protects cells against galectin-3-induced apoptosis J. Biol. Chem. 283, 22177-85. PubMedID: 18676377
    19. Fujita, N., Tamura, A., Higashidani, A., Tonozuka, T., Freeze, H.H., Nishikawa, A. (2008) The relative contribution of mannose salvage pathways to glycosylation in PMI-deficient mouse embryonic fibroblast cells FEBS J.  275, 788-98. PubMedID: 18215164
    20. Wagner-Rousset, E., Bednarczyk, A., Bussat, M.C., Colas, O., Corvaïa, N., Schaeffer, C., Van Dorsselaer, A., Beck, A. (2008) The way forward, enhanced characterization of therapeutic antibody glycosylation: comparison of three level mass spectrometry-based strategies J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 872, 23-37. PubMedID: 18672411
    21. Graham, D.R., Mitsak, M.J., Elliott, S.T., Chen, D., Whelan, S.A., Hart, G.W., Van Eyk, J.E. (2008) Two-dimensional gel-based approaches for the assessment of N-Linked and O-GlcNAc glycosylation in human and simian immunodeficiency viruses Proteomics 8, 4919-30. PubMedID: 19072736
    22. Park, Y., Zhang, Z., Laremore, T.N., Li, B., Sim, J.S., Im, A.R., Ahn, M.Y., Kim, Y.S., Linhardt, R.J., Glycoconj, J. (2008) Variation of acharan sulfate and monosaccharide composition and analysis of neutral N-glycans in African giant snail (Achatina fulica) Glycoconjugate Journal 25, 863-77. PubMedID: 18670878
    23. Rasmussen, T.N., Plenge, P., Bay, T., Egebjerg, J., Gether, U. (2009) A single nucleotide polymorphism in the human serotonin transporter introduces a new site for N-linked glycosylation Neuropharmacology 57, 287-94. PubMedID: 19500602
    24. Gong, B., Cukan, M., Fisher, R., Li, H., Stadheim, T.A., Gerngross, T. (2009) Characterization of N-linked glycosylation on recombinant glycoproteins produced in Pichia pastoris using ESI-MS and MALDI-TOF Methods Mol. Biol. 534, 213-23.
    25. Gefter, J.V., Shaufl, A.L., Fink, M.P., Delude, R.L. (2009) Comparison of distinct protein isoforms of the receptor for advanced glycation end-products expressed in murine tissues and cell lines Cell Tissue Res. 337, 79-89. PubMedID: 19415334
    26. Lee, K.J., Jung, J.H., Lee, J.M., So, Y., Kwon, O., Callewaert, N., Kang, H.A., Ko, K., Oh, D.B. (2009) High-throughput quantitative analysis of plant N-glycan using a DNA sequencer Biochem. Biophys. Res. Commun. 380, 223-9. PubMedID: 19167352
    27. Lee, K.J., Jung, J.H., Lee, J.M,, So, Y., Kwon, O., Callewaert, N., Kang, H.A., Ko, K., Oh, D.B. (2009) High-throughput quantitative analysis of plant N-glycan using a DNA sequencer Biochem. Biophys. Res. Commun.  380, 223-9. PubMedID: 19167352
    28. Piro, J.R., Harris, B.T., Nishina, K., Soto, C., Morales, R., Rees, J.R., Supattapone, S. (2009) Prion protein glycosylation is not required for strain-specific neurotropism  J Virol.  83, 5321-8.
    29. Boeggeman, E., Ramakrishnan, B., Pasek, M., Manzoni, M., Puri, A., Loomis, K.H., Waybright, T.J., Qasba, P.K. (2009) Site specific conjugation of fluoroprobes to the remodeled Fc N-glycans of monoclonal antibodies using mutant glycosyltransferases: application for cell surface antigen detection Bioconjug. Chem. 20, 1228-36. PubMedID: 19425533
    30. Velho, A.M., Jarvis, S.M. (2009) Topological studies of hSVCT1, the human sodium-dependent vitamin C transporter and the influence of N-glycosylation on its intracellular targeting Exp. Cell Res. 315, 2312-21. PubMedID: 19379732
    31. Magnelli PE, Bielik AM, Guthrie EP. (2011) Identification and Characterization of Protein Glycosylation using Specific Endo- and Exoglycosidases J Vis Exp. 58, pii: 3749. PubMedID: 21987255, DOI: 10.3791/3749
    32. Magnelli P, Bielik A, Guthrie E.  (2012) Identification and Characterization of Protein Methods Mol Biol. 801, 189-211.