Novel sialic acid derivatives lock open the 150-loop of an influenza A virus group-1 sialidase

Rudrawar, Santosh; Dyason, Jeffrey Clifford; Rameix‐Welti, Marie‐Anne; Rose, Faith J.; Kerry, Philip S.; Russell, Rosemary; Werf, Sylvie van der; Thomson, Robin Joy; Naffakh, Nadia; Itzstein, Mark von

doi:10.1038/ncomms1114

Cited by 106 publications

(112 citation statements)

References 32 publications

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“…Yet, group 1-specific inhibitors like 3-(p-tolyl)allylNeu5Ac2en also inhibit 09N1 with an affinity similar to that of other group 1 NAs (22). Therefore, we speculate that the atypical group 1 09N1 may possess a 150-loop that is more easily opened than those of group 2 NAs.…”

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confidence: 82%

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Structure of Influenza Virus N7: the Last Piece of the Neuraminidase “Jigsaw” Puzzle

Sun

et al. 2014

J Virol

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There are nine subtypes of influenza A virus neuraminidase (NA), N1 to N9. In addition, influenza B virus also contains NA, and there are two influenza virus NA-like molecules, N10 and N11, which were recently identified from bats. Crystal structures for all of these proteins have been solved, with the exception of N7, and there is no published report of N6, although a structure has been deposited in the Protein Data Bank. Here, we present the N7 and N6 structures at 2.1 Å and 1.8 Å, respectively. Structural comparison of all NA subtypes shows that both N7 and N6 highly resemble typical group 2 NA structures with some special characteristics, including an additional cavity adjacent to their active sites formed by novel 340-loop conformations. Comparative analysis also revealed new structural insights into the N-glycosylation, calcium binding, and second sialic acid binding site of influenza virus NA. This comprehensive study is critical for understanding the complexity of the most successful influenza drug target and for the structure-based design of novel influenza inhibitors. IMPORTANCE Influenza viruses impose a great burden on society, by the human-adapted seasonal types as well as by variants that occasionallyjump from the avian reservoir to infect humans. The surface glycoprotein neuraminidase (NA) is essential for the propagation of the virus and currently the most successfully drug-targeted molecule. Therefore, the structural and functional analysis of NA is critical for the prevention and control of influenza infections. There are nine subtypes of influenza A virus NA (N1 to N9). In addition, influenza B virus also contains NA, and there are two influenza NA-like molecules, N10 and N11, which were recently identified in bats. Crystal structures for all of these proteins have been solved and reported with the exception of N7 and N6. The structural analysis of influenza virus N7 and N6 presented in this study therefore completes the puzzle and adds to a comprehensive understanding of influenza virus NA.

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confidence: 82%

“…This cavity is formed by the 150-loop and was therefore named the 150-cavity. Novel inhibitors, including 3-(p-tolyl)allyl-Neu5Ac2en, have been successfully designed to target this cavity, thereby conferring selectivity toward group 1 NAs (22).…”

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confidence: 99%

Structure of Influenza Virus N7: the Last Piece of the Neuraminidase “Jigsaw” Puzzle

Sun

et al. 2014

J Virol

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“…Infectious titers (doses infecting 50% of the cell culture [TCID 50 ]/50 l) were performed in confluent MDCK and MDCK-SIAT1 cells as previously described (42). The cytopathic effect was visible after incubation at 34°C for 96 h, and the test was revealed by hemagglutination tests performed with 0.5% chicken red blood.…”

Section: Methodsmentioning

confidence: 99%

Combinatorial Effect of Two Framework Mutations (E119V and I222L) in the Neuraminidase Active Site of H3N2 Influenza Virus on Resistance to Oseltamivir

Richard

Ferraris

Erny

et al. 2011

Antimicrob Agents Chemother

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Neuraminidase (NA) inhibitors (NIs) are the first line of defense against influenza virus. Reverse genetics experiments allow the study of resistance mechanisms by anticipating the impacts of mutations to the virus. To look at the possibility of an increased effect on the resistance phenotype of a combination of framework mutations, known to confer resistance to oseltamivir or zanamivir, with limited effect on virus fitness, we constructed 4 viruses by reverse genetics in the A/Moscow/10/99 H3N2 background containing double mutations in their neuraminidase genes: E119D؉I222L, E119V؉I222L, D198N؉I222L, and H274Y؉I222L (N2 numbering). Among the viruses produced, the E119D؉I222L mutant virus was not able to grow without bacterial NA complementation and the D198N؉I222L mutant and H274Y؉I222L mutant were not stable after passages in MDCK cells. The E119V؉I222L mutant was stable after five passages in MDCK cells. This E119V-and-I222L combination had a combinatorial effect on oseltamivir resistance. The total NA activity of the E119V؉I222L mutant was low (5% compared to that of the wild-type virus). This drop in NA activity resulted from a decreased NA quantity in the virion in comparison to that of the wild-type virus (1.4% of that of the wild type). In MDCK-SIAT1 cells, the E119V؉I222L mutant virus did not present a replicative advantage over the wild-type virus, even in the presence of oseltamivir. Double mutations combining two framework mutations in the NA gene still have to be monitored, as they could induce a high level of resistance to NIs, without impairing the NA affinity. Our study allows a better understanding of the diversity of the mechanisms of resistance to NIs.The influenza A virus presents two major surface glycoproteins: hemagglutinin (HA) and neuraminidase (NA). HA mediates virus entry into the cell by binding to terminal sialic acid (N-acetyl neuraminic acid) of cellular glycoconjugates (52). NA, through its sialidase activity, facilitates the elution of progeny virions from infected cells through the cleavage of terminal sialic acid from viral and cellular glycoconjugates (36,44,45).Among the nine NA subtypes described, the crystal structures of the N9 (5, 57), N2 (58, 59), N6, and N1/N4/N8 subtypes (51) have been resolved. The NA head is a tetramer composed of four identical subunits arranged with a circular 4-fold symmetry. Each monomer contains six beta sheets composed of 4 antiparallel strands. The NA active site is located centrally on each unit and forms a pocket composed of conserved residues found in all influenza A virus NA (16). All these residues form the wall of the catalytic pocket. It includes catalytic sites (R118, D151, R152, R224, E276, R292, R371, and Y406) that interact directly with the substrate, sialic acid, and framework sites (E119, R156, W178, S179, D/N198, I222, E227, H274, E277, N294, and E425) that interact with catalytic residues to stabilize the active site (15).Due to its conserved structure and its essential role in viral replication, NA is an attractive antiviral t...

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“…Extensive structural and functional studies of group 2 influenza NAs have led to the development of NA as the most successful drug target against flu to date (10-16). The recent identification of group 1 NA structures, including H5N1 NA (17), pandemic 2009 H1N1 NA (09N1) (18), 1918 H1N1 NA (18N1) (19), N4 (17), N5 (20), and N8 (17), further illustrates the complexity of influenza NA structures and offers some ideas for the design of next-generation NA inhibitors (21,22). However, the identification of N10 revealed an NA-like protein that is significantly divergent from other influenza NAs (7), and thus whether N10 has special structural and/or functional features remains unknown.…”

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confidence: 99%

Structural and functional characterization of neuraminidase-like molecule N10 derived from bat influenza A virus

Sun

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

108

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The recent discovery of the unique genome of influenza virus H17N10 in bats raises considerable doubt about the origin and evolution of influenza A viruses. It also identifies a neuraminidase (NA)-like protein, N10, that is highly divergent from the nine other well-established serotypes of influenza A NA (N1–N9). The structural elucidation and functional characterization of influenza NAs have illustrated the complexity of NA structures, thus raising a key question as to whether N10 has a special structure and function. Here the crystal structure of N10, derived from influenza virus A/little yellow-shouldered bat/Guatemala/153/2009 (H17N10), was solved at a resolution of 2.20 Å. Overall, the structure of N10 was found to be similar to that of the other known influenza NA structures. In vitro enzymatic assays demonstrated that N10 lacks canonical NA activity. A detailed structural analysis revealed dramatic alterations of the conserved active site residues that are unfavorable for the binding and cleavage of terminally linked sialic acid receptors. Furthermore, an unusual 150-loop (residues 147–152) was observed to participate in the intermolecular polar interactions between adjacent N10 molecules of the N10 tetramer. Our study of influenza N10 provides insight into the structure and function of the sialidase superfamily and sheds light on the molecular mechanism of bat influenza virus infection.

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Novel sialic acid derivatives lock open the 150-loop of an influenza A virus group-1 sialidase

Cited by 106 publications

References 32 publications

Structure of Influenza Virus N7: the Last Piece of the Neuraminidase “Jigsaw” Puzzle

Structure of Influenza Virus N7: the Last Piece of the Neuraminidase “Jigsaw” Puzzle

Combinatorial Effect of Two Framework Mutations (E119V and I222L) in the Neuraminidase Active Site of H3N2 Influenza Virus on Resistance to Oseltamivir

Structural and functional characterization of neuraminidase-like molecule N10 derived from bat influenza A virus

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