glyco application

Glycobiology & Proteomics

Glycobiology is the study of the structure, function and biology of carbohydrates, often called glycans, which are widely distributed in nature (1). It is a small but rapidly growing field in biology, with relevance to biomedicine, biotechnology, biofuels and basic research. In eukaryotic cells the majority of proteins are post-translationally modified (2). A common modification, essential for cell viability, is the attachment of glycans. For a visual representation, see the N-linked and O-linked glycosylation tab below.

Glycans define many properties of glycoconjugates (glycoproteins and glycolipids). For instance, it is largely through glycan–protein interactions that cell–cell and cell–pathogen contacts occur. Likewise, glycan molecules modulate many other processes important for cell and tissue differentiation, metabolic and gene regulation, protein activity, protein clearance, transport, and more (3-10). For more information, see the role of carbohydrates in the inflammation response tab below.

Using Glycosidases to Remove, Trim, or Modify Glycans on Therapeutic Proteins 

 
  1. Spiro, R. G. (2002) Glycobiology 12, 43R-56R. PMID: 112042244
  2. Khoury G.A. et al (2011) Scientific Reports 1: 90. PMID: 22034591
  3. Varki A. (1993) Glycobiology. 3(2):97-130. PMID: 8490246
  4. Zhao Y.Y. et al (2008) Cancer Sci. 99(7):1304-10. PMID: 18492092
  5. Zhao Y. et al (2008) FEBS J. 275(9):1939-48. PMID: 18384383
  6. Skropeta D. (2009) Bioorg Med Chem. 17(7):2645-53. PMID: 19285412
  7. Neu U. et al (2011) Curr Opin Struct Biol. 21(5):610-8. PMID: 21917445
  8. Cerliani J.P. et al (2011) J Clin Immunol. 31(1):10-21. PMID: 21184154
  9. Aarnoudse C.A. et al (2006) Curr Opin Immunol. 18(1):105-11. PMID: 16303292
  10. Arnold J.N. (2006) Immunol Lett. 106(2):103-10. PMID: 16814399

Glycobiology & Proteomics includes these areas of focus:

Sequencing Glycans
Biosynthesis of Glycans in Eukaryotes
Removal of N-Linked & O-Linked Glycans from Glycoproteins
Glycoprotein Production in Various Expression Systems
Depolymerization of Heparin/HS
MS Analysis of GAGs

FAQs for Glycobiology & Proteomics

    Publications related to Glycobiology & Proteomics

  1. Kwon HM, Lee KH, Han BW, Han MR, Kim DH, Kim DE 2014. An RNA aptamer that specifically binds to the glycosylated hemagglutinin of avian influenza virus and suppresses viral infection in cells PLoS One. 9(5), PubMedID: 24835440, DOI: 10.1371/journal.pone.0097574
  2. Stech M, Quast RB, Sachse R, Schulze C, Wüstenhagen DA, Kubick S 2014. A continuous-exchange cell-free protein synthesis system based on extracts from cultured insect cells PLoS One. 9(5), PubMedID: 24804975, DOI: 10.1371/journal.pone.0096635
  3. Botto L, Cunati D, Coco S, Sesana S, Bulbarelli A, Biasini E, Colombo L, Negro A, Chiesa R, Masserini M, Palestini P 2014. Role of lipid rafts and GM1 in the segregation and processing of prion protein PLoS One. 9(5), PubMedID: 24859148, DOI: 10.1371/journal.pone.0098344
  4. Rosenbaum EE, Vasiljevic E, Brehm KS, Colley NJ 2014. Mutations in four glycosyl hydrolases reveal a highly coordinated pathway for rhodopsin biosynthesis and N-glycan trimming in Drosophila melanogaster PLoS Genet. 10(5), PubMedID: 24785692, DOI: 10.1371/journal.pgen.1004349
  5. Rosenbaek LL, Kortenoeven ML, Aroankins TS, Fenton RA 2014. Phosphorylation decreases ubiquitylation of the thiazide-sensitive cotransporter NCC and subsequent clathrin-mediated endocytosis J Biol Chem. 289(19), PubMedID: 24668812, DOI: 10.1074/jbc.M113.543710
  6. Möykkynen T, Coleman SK, Semenov A, Keinänen K 2014. The N-terminal domain modulates α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor desensitization J Biol Chem. 289(19), PubMedID: 24652293, DOI: 10.1074/jbc.M113.526301
  7. Arakel EC, Brandenburg S, Uchida K, Zhang H, Lin YW, Kohl T, Schrul B, Sulkin MS, Efimov IR, Nichols CG, Lehnart SE, Schwappach B 2014. Tuning the electrical properties of the heart by differential trafficking of KATP ion channel complexes J Cell Sci. 127(Pt 9), PubMedID: 24569881, DOI: 10.1242/jcs.141440
  8. Itahana Y, Han R, Barbier S, Lei Z, Rozen S, Itahana K 2014. The uric acid transporter SLC2A9 is a direct target gene of the tumor suppressor p53 contributing to antioxidant defense Oncogene. , PubMedID: 24858040, DOI: 10.1038/onc.2014.119
  9. Wicht O, Burkard C, de Haan CA, van Kuppeveld FJ, Rottier PJ, Bosch BJ 2014. Identification and Characterization of a Proteolytically Primed Form of the Murine Coronavirus Spike Proteins after Fusion with the Target Cell J Virol. 88(9), PubMedID: 24554652, DOI: 10.1128/JVI.03451-13
  10. Wright CR, Brown EL, Della-Gatta PA, Ward AC, Lynch GS, Russell AP 2014. G-CSF does not influence C2C12 myogenesis despite receptor expression in healthy and dystrophic skeletal muscle Front Physiol. 5, PubMedID: 24822049, DOI: 10.3389/fphys.2014.00170
  11. Zhao H, Blazanovic K, Choi Y, Bailey-Kellogg C, Griswold KE 2014. Gene and protein sequence optimization for high-level production of fully active and aglycosylated lysostaphin in Pichia pastoris Appl Environ Microbiol. 80(9), PubMedID: 24561590, DOI: 10.1128/AEM.03914-13
  12. Haller G, Li P, Esch C, Hsu S, Goate AM, Steinbach JH 2014. Functional characterization improves associations between rare non-synonymous variants in CHRNB4 and smoking behavior PLoS One. 9(5), PubMedID: 24804708, DOI: 10.1371/journal.pone.0096753
  13. 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 , PubMedID: 19379732, DOI:
  14. 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 , PubMedID: 19425533, DOI:
  15. 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 , PubMedID: , DOI:
  16. 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 , PubMedID: 19500602, DOI:
  17. 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 , PubMedID: 19415334, DOI:
  18. 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, PubMedID: 18672411, DOI:
  19. 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. 872, PubMedID: 19072736, DOI:
  20. Wong-Madden, S.T., Landry, D., and Guthrie, E.P. 1997. Discovery and Uses of Novel Glycosidases Techniques in Glycobiology. , PubMedID: , DOI:

N-Linked and O-Linked Glycosylation

N-linked glycosylation occurs through consensus asparagine residues of the protein, while O-glycosylation occurs through serine or threonine residues.

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