Trisaccharide containing α2,3-linked sialic acid is a receptor for mumps virus.
Mumps virus (MuV) remains an important pathogen worldwide, causing epidemic parotitis, orchitis, meningitis, and encephalitis. Here we show that MuV preferentially uses a trisaccharide containing α2,3-linked sialic acid in unbranched sugar chains as a receptor. Crystal structures of the MuV attachment protein hemagglutinin-neuraminidase (MuV-HN) alone and in complex with the α2,3-sialylated trisaccharide revealed that in addition to the interaction between the MuV-HN active site residues and sialic acid, other residues, including an aromatic residue, stabilize the third sugar of the trisaccharide. The importance of the aromatic residue and the third sugar in the MuV-HN-receptor interaction was confirmed by computational energy calculations, isothermal titration calorimetry studies, and glycan-binding assays. Furthermore, MuV-HN was found to bind more efficiently to unbranched α2,3-sialylated sugar chains compared with branched ones. Importantly, the strategically located aromatic residue is conserved among the HN proteins of sialic acid-using paramyxoviruses, and alanine substitution compromised their ability to support cell-cell fusion. These results suggest that not only the terminal sialic acid but also the adjacent sugar moiety contribute to receptor function for mumps and these paramyxoviruses. The distribution of structurally different sialylated glycans in tissues and organs may explain in part MuV's distinct tropism to glandular tissues and the central nervous system. In the crystal structure, the epitopes for neutralizing antibodies are located around the α-helices of MuV-HN that are not well conserved in amino acid sequences among different genotypes of MuV. This may explain the fact that MuV reinfection sometimes occurs.
Department of Virology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan.
Marie
M
Kubota
Laboratory of Environmental Microbiology, Faculty of Medicine, University of Tsukuba, Tsukuba 305-8575, Japan; ktakeuch@md.tsukuba.ac.jp takaoh@virology.med.kyushu-u.ac.jp.
Kaoru
K
Takeuchi
Department of Virology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan.
Shumpei
S
Watanabe
Department of Virology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan.
Shinji
S
Ohno
Division of Structural Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka 812-8582, Japan.
Rei
R
Matsuoka
Division of Structural Biology, Medical Institute of Bioregulation, Kyushu University, Fukuoka 812-8582, Japan.
Daisuke
D
Kohda
Department of Functional Glycomics, Life Science Research Center, Kagawa University, Kagawa 761-0793, Japan.
Shin-Ichi
SI
Nakakita
Health Science Hills, College of Life and Health Sciences, Chubu University, Kasugai 487-8501, Japan.
Hiroaki
H
Hiramatsu
Health Science Hills, College of Life and Health Sciences, Chubu University, Kasugai 487-8501, Japan.
Yasuo
Y
Suzuki
Laboratory of Viral Infection, Kitasato Institute for Life Sciences, Tokyo 108-8641, Japan.
Tetsuo
T
Nakayama
Department of Biotechnology and Agricultural Bioinformatics Research Unit, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
Tohru
T
Terada
Department of Biotechnology and Agricultural Bioinformatics Research Unit, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
Kentaro
K
Shimizu
Photon Factory, Institute of Materials Structure Science, High-Energy Accelerator Research Organization, Tsukuba 305-0801, Japan.
Nobutaka
N
Shimizu
Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka 812-8582, Japan.
Mitsunori
M
Shiroishi
Department of Virology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan.
Yusuke
Y
Yanagi
Department of Virology, Faculty of Medicine, Kyushu University, Fukuoka 812-8582, Japan; ktakeuch@md.tsukuba.ac.jp takaoh@virology.med.kyushu-u.ac.jp.
Takao
T
Hashiguchi
D000818:Animals; D057134:Antibodies, Neutralizing / Q000737:chemistry; D002459:Cell Fusion; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D000939:Epitopes / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D007785:Lactose / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D009109:Mumps virus / Q000378:metabolism*; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D019158:N-Acetylneuraminic Acid / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D000072417:Protein Domains; D011991:Receptors, Virus / Q000737:chemistry / Q000378:metabolism*; D013816:Thermodynamics; D014312:Trisaccharides / Q000737:chemistry* / Q000378:metabolism*; D014709:Vero Cells; D014764:Viral Proteins / Q000737:chemistry / Q000378:metabolism
D057134:Antibodies, Neutralizing; D000939:Epitopes; D050505:Mutant Proteins; D011991:Receptors, Virus; D014312:Trisaccharides; D014764:Viral Proteins; D019158:N-Acetylneuraminic Acid; D007785:Lactose
entry; infection; paramyxovirus; receptor; structure
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