A Secondary Sialic Acid Binding Site on Influenza Virus Neuraminidase: Fact or Fiction?

Lai, Jimmy C. C.; Garcia, Jean‐Michel; Dyason, Jeffrey Clifford; Böhm, Raphael; Madge, Paul D.; Rose, Faith J.; Nicholls, John M.; Peiris, Malik; Haselhorst, Thomas; Itzstein, Mark von

doi:10.1002/anie.201108245

Cited by 30 publications

(43 citation statements)

References 11 publications

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“…Of broader relevance, they provide a general paradigm for dynamic, catalysis-driven virus-sialoglycan interactions in relation to host selectivity. This notion is supported by observations for influenza A viruses, where neuraminidase (NA) activity toward multivalent substrates varies with differences in the length of the stalk domain and positively correlates with NA size (Castrucci and Kawaoka, 1993;Els et al, 1985), where NA catalytic activity is modulated by the absence or presence of a second Sia-binding site (Uhlendorff et al, 2009;Varghese et al, 1997) and where changes in NA stalk length and NA lectin affinity have been implicated in host specificity (Blumenkrantz et al, 2013;Castrucci and Kawaoka, 1993;Lai et al, 2012;Li et al, 2014;Matrosovich et al, 1999;Munier et al, 2010;Uhlendorff et al, 2009;Varghese et al, 1997). These observations can be readily interpreted in the context of our data and model proposed.…”

Section: Resultsmentioning

confidence: 82%

Betacoronavirus Adaptation to Humans Involved Progressive Loss of Hemagglutinin-Esterase Lectin Activity

Bakkers

Lang

Feitsma

et al. 2017

Cell Host & Microbe

147

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Human beta1-coronavirus (β1CoV) OC43 emerged relatively recently through a single zoonotic introduction. Like related animal β1CoVs, OC43 uses 9-O-acetylated sialic acid as receptor determinant. β1CoV receptor binding is typically controlled by attachment/fusion spike protein S and receptor-binding/receptor-destroying hemagglutinin-esterase protein HE. We show that following OC43's introduction into humans, HE-mediated receptor binding was selected against and ultimately lost through progressive accumulation of mutations in the HE lectin domain. Consequently, virion-associated receptor-destroying activity toward multivalent glycoconjugates was reduced and altered such that some clustered receptor populations are no longer cleaved. Loss of HE lectin function was also observed for another respiratory human coronavirus, HKU1. This thus appears to be an adaptation to the sialoglycome of the human respiratory tract and for replication in human airways. The findings suggest that the dynamics of virion-glycan interactions contribute to host tropism. Our observations are relevant also to other human respiratory viruses of zoonotic origin, particularly influenza A virus.

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Section: Resultsmentioning

confidence: 82%

Betacoronavirus Adaptation to Humans Involved Progressive Loss of Hemagglutinin-Esterase Lectin Activity

Bakkers

Lang

Feitsma

et al. 2017

Cell Host & Microbe

147

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“…For N9, X-ray crystallography (27) revealed that the 2nd SIA-binding site is a shallow pocket composed of three surface peptide loops, located adjacent to the catalytic site. The presence of a 2nd SIA-binding site in N1 was also demonstrated using saturation transfer difference (STD) nuclear magnetic resonance (NMR) spectroscopy (28). By both hemadsorption assays and STD NMR, it was shown that N1 and N2 proteins of swine and human IAVs display severely reduced binding compared to their avian counterparts (26,28).…”

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confidence: 97%

“…The presence of a 2nd SIA-binding site in N1 was also demonstrated using saturation transfer difference (STD) nuclear magnetic resonance (NMR) spectroscopy (28). By both hemadsorption assays and STD NMR, it was shown that N1 and N2 proteins of swine and human IAVs display severely reduced binding compared to their avian counterparts (26,28). Sequence analysis indicated that five out of six SIA contact residues in the N9 2nd SIA-binding site are highly conserved across all avian NA genotypes.…”

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confidence: 99%

Mutation of the Second Sialic Acid-Binding Site, Resulting in Reduced Neuraminidase Activity, Preceded the Emergence of H7N9 Influenza A Virus

Dai

McBride

Dortmans

et al. 2017

J Virol

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The emergence of the novel influenza A virus (IAV) H7N9 since 2013 has caused concerns about the ability of the virus to spread between humans. Analysis of the receptor-binding properties of the H7 protein of a human isolate revealed modestly increased binding to α2,6 sialosides and reduced, but still dominant, binding to α2,3-linked sialic acids (SIAs) compared to a closely related avian H7N9 virus from 2008. Here, we show that the corresponding N9 neuraminidases (NAs) display equal enzymatic activities on a soluble monovalent substrate and similar substrate specificities on a glycan array. In contrast, solid-phase activity and binding assays demonstrated reduced specific activity and decreased binding of the novel N9 protein. Mutational analysis showed that these differences resulted from substitution T401A in the 2nd SIA-binding site, indicating that substrate binding via this site enhances NA catalytic activity. Substitution T401A in the novel N9 protein appears to functionally mimic the substitutions that are found in the 2nd SIA-binding site of NA proteins of avian-derived IAVs that became human pandemic viruses. Our phylogenetic analyses show that substitution T401A occurred prior to substitutions in hemagglutinin (HA), causing the altered receptor-binding properties mentioned above. Hence, in contrast to the widespread assumption that such changes in NA are obtained only after acquisition of functional changes in HA, our data indicate that mutations in the 2nd SIA-binding site may have enabled and even driven the acquisition of altered HA receptor-binding properties and may have contributed to the spread of the novel H7N9 viruses. Novel H7N9 IAVs continue to cause human infections and pose an ongoing public health threat. Here, we show that their N9 proteins display reduced binding to and lower enzymatic activity against multivalent substrates, resulting from mutation of the 2nd sialic acid-binding site. This mutation preceded and may have driven the selection of substitutions in H7 that modify H7 receptor-binding properties. Of note, all animal IAVs that managed to cross the host species barrier and became human viruses carry mutated 2nd sialic acid-binding sites. Screening of animal IAVs to monitor their potential to cross the host species barrier should therefore focus not only on the HA protein, but also on the functional properties of NA.

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“…Sialic acids bind to influenza NA at two independent binding sites: the catalytic site and the secondary binding site. With the catalytic site being blocked by oseltamivir carboxylate, STD‐NMR signals of sialyllactoses binding to the secondary site could be evaluated . Differential STD‐NMR spectra were generated by subtracting the secondary site binding signals from the overall NA‐SL binding spectra, which reflect only the binding signals of sialyllactoses to the NA catalytic site.…”

Section: Resultsmentioning

confidence: 99%

Investigation of the binding and cleavage characteristics of N1 neuraminidases from avian, seasonal, and pandemic influenza viruses using saturation transfer difference nuclear magnetic resonance

Garcia

Lai

Haselhorst

et al. 2013

Influenza Resp Viruses

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ObjectivesThe main function of influenza neuraminidase (NA) involves enzymatic cleavage of sialic acid from the surface of host cells resulting in the release of the newly produced virions from infected cells, as well as aiding the movement of virions through sialylated mucus present in the respiratory tract. However, there has previously been little information on the binding affinity of different forms of sialylated glycan with NA. Our objectives were then to investigate both sialic acid binding and cleavage of neuraminidase at an atomic resolution level.DesignNuclear magnetic resonance (NMR) spectroscopy was used to investigate pH and temperature effects on binding and cleavage as well as to interrogate the selectivity of human-like or avian-like receptors for influenza neuraminidase N1 derived from a range of different influenza virus strains including human seasonal H1N1, H1N1pdm09 and avian H5N1.ResultsWe demonstrated that an acidic pH and physiological temperature are required for efficient NA enzymatic activity; however a change in the pH had a minimum effect on the NA-sialic acid binding affinity. Our data comparing α-2,3- and α-2,6-sialyllactose indicated that the variation in neuraminidase activity on different ligands correlated with a change in binding affinity. Epitope mapping of the sialylglycans interacting with NAs from different viral origin showed different binding profiles suggesting that different binding conformations were adopted.ConclusionsThe data presented in this study demonstrated that physicochemical conditions (pH in particular) could affect the NA enzymatic activity with minor effect on ligand binding. NA cleavage specificity seemed to be associated with a difference in binding affinity to different ligands, suggesting a relationship between the two events. These findings have implications regarding the replication cycle of influenza infection in the host where different sialidase activities would influence penetration through the respiratory mucin barrier and the release of the newly generated virus from the infected cells.

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A Secondary Sialic Acid Binding Site on Influenza Virus Neuraminidase: Fact or Fiction?

Cited by 30 publications

References 11 publications

Betacoronavirus Adaptation to Humans Involved Progressive Loss of Hemagglutinin-Esterase Lectin Activity

Betacoronavirus Adaptation to Humans Involved Progressive Loss of Hemagglutinin-Esterase Lectin Activity

Mutation of the Second Sialic Acid-Binding Site, Resulting in Reduced Neuraminidase Activity, Preceded the Emergence of H7N9 Influenza A Virus

Investigation of the binding and cleavage characteristics of N1 neuraminidases from avian, seasonal, and pandemic influenza viruses using saturation transfer difference nuclear magnetic resonance

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