title: Kinetic, thermodynamic and structural analysis of tamiphosphor binding to neuraminidase of H1N1 (2009) pandemic influenza.
abstract: Influenza virus causes severe respiratory infections that are responsible for up to half a million deaths worldwide each year. Two inhibitors targeting viral neuraminidase have been approved to date (oseltamivir, zanamivir). However, the rapid development of antiviral drug resistance and the efficient transmission of resistant viruses among humans represent serious threats to public health. The approved influenza neuraminidase inhibitors have (oxa)cyclohexene scaffolds designed to mimic the oxonium transition state during enzymatic cleavage of sialic acid. Their active forms contain a carboxylate that interacts with three arginine residues in the enzyme active site. Recently, the phosphonate group was successfully used as an isostere of the carboxylate in oseltamivir, and the resulting compound, tamiphosphor, was identified as a highly active neuraminidase inhibitor. However, the structure of the complex of this promising inhibitor with neuraminidase has not yet been reported. Here, we analyzed the interaction of a set of oseltamivir and tamiphosphor derivatives with neuraminidase from the A/California/07/2009 (H1N1) influenza virus. We thermodynamically characterized the binding of oseltamivir carboxylate or tamiphosphor to the neuraminidase catalytic domain by protein microcalorimetry, and we determined crystal structure of the catalytic domain in complex with tamiphosphor at 1.8 Å resolution. This structural information should aid rational design of the next generation of neuraminidase inhibitors.
authors.affiliation: Department of Organic Chemistry, Faculty of Science, Charles University, Hlavova 8, 128 00, Prague 2, Czech Republic.
authors.forename: Carlos Berenguer
authors.initials: CB
authors.lastname: Albiñana
authors.affiliation: Department of Organic Chemistry, Faculty of Science, Charles University, Hlavova 8, 128 00, Prague 2, Czech Republic.
authors.forename: Aleš
authors.initials: A
authors.lastname: Machara
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead Sciences and IOCB Research Center, Flemingovo n. 2, 166 10, Prague 6, Czech Republic; Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídeňská 1083, 140 00, Prague 4, Czech Republic.
authors.forename: Pavlína
authors.initials: P
authors.lastname: Řezáčová
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead Sciences and IOCB Research Center, Flemingovo n. 2, 166 10, Prague 6, Czech Republic.
authors.forename: Petr
authors.initials: P
authors.lastname: Pachl
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead Sciences and IOCB Research Center, Flemingovo n. 2, 166 10, Prague 6, Czech Republic; Department of Biochemistry, Faculty of Science, Charles University, Hlavova 8, 128 00, Prague 2, Czech Republic. Electronic address: konval@uochb.cas.cz.
authors.forename: Jan
authors.initials: J
authors.lastname: Konvalinka
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Gilead Sciences and IOCB Research Center, Flemingovo n. 2, 166 10, Prague 6, Czech Republic. Electronic address: milan.kozisek@uochb.cas.cz.
authors.forename: Milan
authors.initials: M
authors.lastname: Kožíšek
mesh_terms: D000998:Antiviral Agents / Q000737:chemistry* / Q000494:pharmacology; D020134:Catalytic Domain; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000201:enzymology*; D007251:Influenza, Human / Q000821:virology; D007700:Kinetics; D009439:Neuraminidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D053139:Oseltamivir / Q000031:analogs & derivatives* / Q000378:metabolism / Q000627:therapeutic use; D058873:Pandemics; D010757:Phosphorous Acids / Q000378:metabolism* / Q000627:therapeutic use; D011485:Protein Binding; D013816:Thermodynamics
chemical_list: D000998:Antiviral Agents; D004791:Enzyme Inhibitors; D010757:Phosphorous Acids; C530824:tamiphosphor; D053139:Oseltamivir; D009439:Neuraminidase
keywords: Crystal structure; Influenza neuraminidase; Isothermal titration calorimetry; Lattice-translocation defect; Oseltamivir; Tamiphosphor