Identifying Carbohydrate Ligands of a Norovirus P Particle using a Catch and Release Electrospray Ionization Mass Spectrometry Assay
- 561 Downloads
Noroviruses (NoVs), the major cause of epidemic acute gastroenteritis, recognize human histo-blood group antigens (HBGAs), which are present as free oligosaccharides in bodily fluid or glycolipids and glycoproteins on the surfaces of cells. The subviral P particle formed by the protruding (P) domain of the NoV capsid protein serves as a useful model for the study NoV–HBGA interactions. Here, we demonstrate the application of a catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) assay for screening carbohydrate libraries against the P particle to rapidly identify NoV ligands and potential inhibitors. Carbohydrate libraries of 50 and 146 compounds, which included 18 and 24 analogs of HBGA receptors, respectively, were screened against the P particle of VA387, a member of the predominant GII.4 NoVs. Deprotonated ions corresponding to the P particle bound to carbohydrates were isolated and subjected to collision-induced dissociation to release the ligands in their deprotonated forms. The released ligands were identified by ion mobility separation followed by mass analysis. All 13 and 16 HBGA ligands with intrinsic affinities >500 M–1 were identified in the 50 and the 146 compound libraries, respectively. Furthermore, screening revealed interactions with a series of oligosaccharides with structures found in the cell wall of mycobacteria and human milk. The affinities of these newly discovered ligands are comparable to those of the HBGA receptors, as estimated from the relative abundance of released ligand ions.
Key wordsCatch and release Electrospray ionization mass spectrometry Norovirus P particle Carbohydrate
The authors thank the Alberta Glycomics Centre and the National Institutes of Health of the Unites States of America for supporting this research. They thank Professors T. Lowary (University of Alberta) and C.-C. Ling (University of Calgary), as well as Alberta Innovates Technology Futures for generously providing some of the carbohydrates used in this work. L.H. also acknowledges an Alberta Innovates Graduate Student Scholarship.
- 3.Huang, P., Farkas, T., Marionneau, S., Zhong, W., Ruvoen-Clouet, N., Morrow, A.L., Altaye, M., Pickering, L.K., Newburg, D.S., LePendu, J., Jiang, X.: Noroviruses bind to human ABO, Lewis, and secretor histo-blood group antigens: Identification of 4 distinct strain-specific patterns. J. Infect. Dis 188, 19–31 (2003)CrossRefGoogle Scholar
- 4.Huang, P., Farkas, T., Zhong, W., Thornton, S., Morrow, A.L., Jiang, X.: Norovirus and histo-blood group antigens: demonstration of a wide spectrum of strain specificities and classification of two major binding groups among multiple binding patterns. J. Virol 79, 6714–6722 (2005)CrossRefGoogle Scholar
- 16.Jiang, X., Wang, M., Graham, D.Y., Estes, M.K.: Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein. J. Virol 66, 6527–6532 (1992)Google Scholar
- 20.Bu, W., Mamedova, A., Tan, M., Xia, M., Jiang, X., Hegde, R.S.: Structural basis for the receptor binding specificity of Norwalk virus. J. Virol. 82, 5340–5347 (2008) native mass spectrometry. J. Struct. Biol. 177, 273–282 (2012)Google Scholar
- 24.Hansman, G.S., Biertumpfel, C., Georgiev, I., McLellan, J.S., Chen, L., Zhou, T.Q., Katayama, K., Kwong, P.D.: Crystal structures of GII.10 and GII.12 norovirus protruding domains in complex with histo-blood group antigens reveal details for a potential site of vulnerability. J. Virol 85, 6687–6701 (2011)CrossRefGoogle Scholar
- 44.Cederkvist, F., Zamfir, A.D., Bahrke, S., Eijsink, V.G.H., Sorlie, M., Peter-Katalinic, J., Peter, M.G.: Identification of a high-affinity-binding oligosaccharide by (+) nanoelectrospray quadrupole time-of-flight tandem mass spectrometry of a noncovalent enzyme-ligand complex. Angew. Chem. Int. Ed 45, 2429–2434 (2006)CrossRefGoogle Scholar
- 45.Abzalimov, R.R., Dubin, P.L., Kaltashov, I.A.: Glycosaminoglycans as naturally occurring combinatorial libraries: spectrometry-based strategy for characterization of anti-thrombin interaction strategy with low molecular weight heparin and heparin oligomers. Anal. Chem 79, 6055–6063 (2007)CrossRefGoogle Scholar