Abstract
Recently, we reported the characterization of the glycans attached at the 11 N-glycosylation sites of Hepatitis C virus E2 envelope glycoprotein by tandem mass spectrometry. Infections caused by Hepatitis C virus represent the main cause of liver diseases such as hepatitis, cirrhosis and hepatocellular carcinoma. The N-linked sugars consist primarily of high mannose glycans, with structures ranging from the minimal core structure, Man3GlcNAc2 (Man3) up to 12 hexose residues attached to the GlcNAc-ß(l–4)-GlcNAc core (depicted as Hex3Man9GlcNAc2). Furthermore, the site N41 (N423) was observed to contain complex type glycans with the structures Man3-GlcNAc and Man3-GlcNAcFuc, in addition to the high mannose population Man3 through Man6, while the site N48 (N430) was occupied exclusively with complex type glycans (Man3-Fuc, Man3-GlcNAcFuc and Man3-GlcNAc2Fuc). The present contribution summarizes our experimental observations upon the factors which may have an impact on the CID tandem mass spectra of glycopeptides.
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References
Marsh, M. and A. Helenius, Virus entry: open sesame. Cell, 2006. 124(4): 729-40.
Slawson, C., M.P. Housley, and G.W. Hart, O-GlcNAc cycling: how a single sugar post- translational modification is changing the way we think about signaling networks. J Cell Biochem, 2006. 97(1): 71-83.
Smith, A.E. and A. Helenius, How viruses enter animal cells. Science, 2004. 304(5668): 237-42.
Wormald, M.R. and R.A. Dwek, Glycoproteins: glycan presentation and protein-fold stabi- lity. Structure, 1999. 7(7): R155-60.
Apweiler, R., H. Hermjakob, and N. Sharon, On the frequency of protein glycosylation, as deduced from analysis of the SWISS-PROT database. Biochim Biophys Acta, 1999. 1473(1): 4-8.
Kornfeld, R. and S. Kornfeld, Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem, 1985. 54: 631-64.
Gavel, Y. and G. von Heijne, Sequence differences between glycosylated and non- glycosylated Asn-X-Thr/Ser acceptor sites: implications for protein engineering. Protein Eng, 1990. 3(5): 433-42.
Srebalus Barnes, C.A. and A. Lim, Applications of mass spectrometry for the structural characterization of recombinant protein pharmaceuticals. Mass Spectrom Rev, 2007. 26(3): 370-88.
Fenn, J.B., et al., Electrospray ionization for mass spectrometry of large biomolecules. Sci- ence, 1989. 246(4926): 64-71.
Hillenkamp, F., et al., Matrix-assisted laser desorption/ionization mass spectrometry of bio- polymers. Anal Chem, 1991. 63(24): 1193A-1203A.
Wuhrer, M., A.M. Deelder, and C.H. Hokke, Protein glycosylation analysis by liquid chromatography-mass spectrometry. J Chromatogr B Analyt Technol Biomed Life Sci, 2005.825(2): 124-33.
Zaia, J., Mass spectrometry of oligosaccharides. Mass Spectrom Rev, 2004. 23(3): 161-227.
Huddleston, M.J., M.F. Bean, and S.A. Carr, Collisional fragmentation of glycopeptides by electrospray ionization LC/MS and LC/MS/MS: methods for selective detection of glycopeptides in protein digests. Anal Chem, 1993. 65(7): 877-84.
Ciucanu, I., Per-O-methylation reaction for structural analysis of carbohydrates by mass spectrometry. Anal Chim Acta, 2006. 576(2): 147-55.
Harazono, A., et al., Site-specific N-glycosylation analysis of human plasma ceruloplasmin using liquid chromatography with electrospray ionization tandem mass spectrometry. Anal Biochem, 2006. 348(2): 259-68.
Wuhrer, M., et al., IPSE/alpha-1, a major secretory glycoprotein antigen from schistosome eggs, expresses the Lewis X motif on core-difucosylated N-glycans. Febs J, 2006. 273(10): 2276-92.
Wuhrer, M., et al., Glycoproteomics based on tandem mass spectrometry of glycopeptides. J Chromatogr B Analyt Technol Biomed Life Sci, 2007. 849(1-2): 115-28.
Iacob, R., Perdivara, I., Przybylski, M., and Tomer, K.B., Mass spectrometric characteri-zation of glycosylation of Hepatitis C virus E2 envelope glycoprotein reveals extended micro-heterogeneity of N-glycans. JASMS, 2007.
Rehermann, B. and M. Nascimbeni, Immunology of hepatitis B virus and hepatitis C virus infection. Nat Rev Immunol, 2005. 5(3): 215-29.
Randall, G. and C.M. Rice, Hepatitis C virus cell culture replication systems: their potential use for the development of antiviral therapies. Curr Opin Infect Dis, 2001. 14(6): 743-7.
Duvet, S., et al., Hepatitis C virus glycoprotein complex localization in the endoplasmic reticulum involves a determinant for retention and not retrieval. J Biol Chem, 1998. 273 (48): 32088-95.
Domon, B. and Costello, C.E., A Systematic Nomenclature for Carbohydrate Fragmenta-tions in FAB-MS/MS Spectra of Glycoconjugates. Glycoconjugate J, 1988. 5: 397-409.
Paizs, B. and S. Suhai, Fragmentation pathways of protonated peptides. Mass Spectrom Rev, 2005. 24(4): 508-48
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Perdivara, I., Iacob, R.E., Przybylski, M., Tomer, K.B. (2008). Site Specific Identification of N-Linked Glycosylation in Proteins by Liquid Chromatography–Electrospray Ionization Tandem Mass Spectrometry. In: Popescu, C., Zamfir, A.D., Dinca, N. (eds) Applications of Mass Spectrometry in Life Safety. NATO Science for Peace and Security Series A: Chemistry and Biology. Springer, Dordrecht. https://doi.org/10.1007/978-1-4020-8811-7_8
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