Abstract
Many viruses utilize cell-surface glycans as receptors for host cell entry. Viral surface glycoproteins specifically interact with glycan motifs, which strongly contributes to viral tropism. Recently, the interactions between host cell glycan receptors and the mumps virus (MuV) hemagglutinin-neuraminidase (MuV-HN) protein were characterized by determining the co-crystal structure of MuV-HN in complex with glycan receptors. Here, we describe protocols for large-scale expression, purification and crystallization of MuV-HN proteins for structural analyses and glycan-binding assays with the overarching goal of investigating glycan–protein interactions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Rubin SA, Sauder CJ, Carbone KM (2013) Mumps virus. In: Knipe DM, Howley PM, Cohen JI, Griffin DE, Lamb RA, Martin MA, Racaniello VR, Roizman B (eds) Fields virology, 6th edn. Lippincott Williams & Wilkins, Philadelphia, pp 1024–1041
Rubin S, Eckhaus M, Rennick LJ et al (2015) Molecular biology, pathogenesis and pathology of mumps virus. J Pathol 235:242–252
Lamb RA, Parks GD (2013) Paramyxoviridae. In: Knipe DM, Howley PM, Cohen JI, Griffin DE, Lamb RA, Martin MA, Racaniello VR, Roizman B (eds) Fields virology, 6th edn. Lippincott Williams & Wilkins, Philadelphia, pp 957–995
Casasnovas JM (2013) Virus-receptor interactions and receptor-mediated virus entry into host cells. Subcell Biochem 68:441–466
Raman R, Tharakaraman K, Sasisekharan V et al (2016) Glycan-protein interactions in viral pathogenesis. Curr Opin Struct Biol 40:153–162
Thompson AJ, De Vries RP, Paulson JC (2019) Virus recognition of glycan receptors. Curr Opin Virol 34:117–129
Kubota M, Takeuchi K, Watanabe S et al (2016) Trisaccharide containing alpha2,3-linked sialic acid is a receptor for mumps virus. Proc Natl Acad Sci U S A 113:11579–11584
Kubota M, Matsuoka R, Suzuki T et al (2019) Molecular mechanism of the flexible glycan-receptor recognition by mumps virus. J Virol 93:e00344–e00319
Reeves PJ, Callewaert N, Contreras R et al (2002) Structure and function in rhodopsin: high-level expression of rhodopsin with restricted and homogeneous N-glycosylation by a tetracycline-inducible N-acetylglucosaminyltransferase I-negative HEK293S stable mammalian cell line. Proc Natl Acad Sci U S A 99:13419–13424
Hashiguchi T, Kajikawa M, Maita N et al (2007) Crystal structure of measles virus hemagglutinin provides insight into effective vaccines. Proc Natl Acad Sci U S A 104:19535–19540
Rayment I, Rypniewski WR, Schmidt-Base K et al (1993) Three-dimensional structure of myosin subfragment-1: a molecular motor. Science 261:50–58
Walter TS, Meier C, Assenberg R et al (2006) Lysine methylation as a routine rescue strategy for protein crystallization. Structure 14:1617–1622
Hashiguchi T, Ose T, Kubota M et al (2012) Crystallization strategy for the glycoprotein-receptor complex between measles virus hemagglutinin and its cellular receptor SLAM. Protein Pept Lett 19:468–473
Aricescu AR, Assenberg R, Bill RM et al (2006) Eukaryotic expression: developments for structural proteomics. Acta Crystallogr D 62:1114–1124
Hashiguchi T, Kajikawa M, Maita N et al (2008) Homogeneous sugar modification improves crystallization of measles virus hemagglutinin. J Virol Methods 149:171–174
Hashiguchi T, Ose T, Kubota M et al (2011) Structure of the measles virus hemagglutinin bound to its cellular receptor SLAM. Nat Struct Mol Biol 18:135–141
Acknowledgments
We thank Kaoru Takeuchi for materials. This study was supported by grants from AMED J-PRIDE (JP19fm0208022h) and the Terumo Foundation for Life Sciences and Arts to T.H.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Kubota, M., Hashiguchi, T. (2020). Large-Scale Expression and Purification of Mumps Virus Hemagglutinin-Neuraminidase for Structural Analyses and Glycan-Binding Assays. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_55
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0430-4_55
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0429-8
Online ISBN: 978-1-0716-0430-4
eBook Packages: Springer Protocols