Measurement of Enzymatic Activity and Specificity of Human and Avian Influenza Neuraminidases from Whole Virus by Glycoarray and MALDI-TOF Mass Spectrometry

Authors

  • Dr. Gwladys Pourceau,

    1. IBMM UMR 5247 CNRS UM1 UM2, Université de Montpellier 2, Place Eugène Bataillon, 34095 Montpellier Cedex 5 (France)
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  • Dr. Yann Chevolot,

    Corresponding author
    1. Université de Lyon, Institut des Nanotechnologies de Lyon, INL, UMR CNRS 5270, site Ecole Centrale de Lyon, 36, Avenue Guy de Collongue, 69134 Ecully Cedex (France)
    • Université de Lyon, Institut des Nanotechnologies de Lyon, INL, UMR CNRS 5270, site Ecole Centrale de Lyon, 36, Avenue Guy de Collongue, 69134 Ecully Cedex (France)
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  • Alice Goudot,

    1. Université de Lyon, Institut des Nanotechnologies de Lyon, INL, UMR CNRS 5270, site Ecole Centrale de Lyon, 36, Avenue Guy de Collongue, 69134 Ecully Cedex (France)
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  • Fabienne Giroux,

    1. Virologie et Pathologie Humaine, CNRS FRE 3011, Université de Lyon 1, Faculté de médecine RTH Laennec, 7, Rue Guillaume Paradin, 69372 Lyon Cedex 08 (France)
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  • Albert Meyer,

    1. IBMM UMR 5247 CNRS UM1 UM2, Université de Montpellier 2, Place Eugène Bataillon, 34095 Montpellier Cedex 5 (France)
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  • Dr. Vincent Moulés,

    1. Virologie et Pathologie Humaine, CNRS FRE 3011, Université de Lyon 1, Faculté de médecine RTH Laennec, 7, Rue Guillaume Paradin, 69372 Lyon Cedex 08 (France)
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  • Prof. Bruno Lina,

    1. Virologie et Pathologie Humaine, CNRS FRE 3011, Université de Lyon 1, Faculté de médecine RTH Laennec, 7, Rue Guillaume Paradin, 69372 Lyon Cedex 08 (France)
    2. Laboratoire de Virologie, Centre de Biologie et de Pathologie Est, Hospices Civils de Lyon, 59, Boulevard Pinel, 69677 Bron Cedex (France)
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  • Dr. Samy Cecioni,

    1. ICBMS UMR 5246 Laboratoire de Chimie Organique 2 Glycochimie, CNRS Université Lyon 1, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne (France)
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  • Dr. Sébastien Vidal,

    1. ICBMS UMR 5246 Laboratoire de Chimie Organique 2 Glycochimie, CNRS Université Lyon 1, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne (France)
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  • Dr. Hai Yu,

    1. Department of Chemistry, University of California, Davis, CA 95616 (USA)
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  • Prof. Xi Chen,

    1. Department of Chemistry, University of California, Davis, CA 95616 (USA)
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  • Dr. Olivier Ferraris,

    1. Virologie et Pathologie Humaine, CNRS FRE 3011, Université de Lyon 1, Faculté de médecine RTH Laennec, 7, Rue Guillaume Paradin, 69372 Lyon Cedex 08 (France)
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  • Dr. Jean-Pierre Praly,

    1. ICBMS UMR 5246 Laboratoire de Chimie Organique 2 Glycochimie, CNRS Université Lyon 1, 43 Boulevard du 11 Novembre 1918, 69622 Villeurbanne (France)
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  • Dr. Eliane Souteyrand,

    1. Université de Lyon, Institut des Nanotechnologies de Lyon, INL, UMR CNRS 5270, site Ecole Centrale de Lyon, 36, Avenue Guy de Collongue, 69134 Ecully Cedex (France)
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  • Dr. Jean-Jacques Vasseur,

    1. IBMM UMR 5247 CNRS UM1 UM2, Université de Montpellier 2, Place Eugène Bataillon, 34095 Montpellier Cedex 5 (France)
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  • Dr. François Morvan

    Corresponding author
    1. IBMM UMR 5247 CNRS UM1 UM2, Université de Montpellier 2, Place Eugène Bataillon, 34095 Montpellier Cedex 5 (France)
    • IBMM UMR 5247 CNRS UM1 UM2, Université de Montpellier 2, Place Eugène Bataillon, 34095 Montpellier Cedex 5 (France)
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Abstract

Influenza neuraminidases hydrolyze the ketosidic linkage between N-acetylneuraminic acid and its adjacent galactose residue in sialosides. This enzyme is a tetrameric protein that plays a critical role in the release of progeny virions. Several methods have been described for the determination of neuraminidase activity, usually based on colorimetric, fluorescent, or chemiluminescent detection. However, only a few of these tests allow discrimination of the sialyl-linkage specificity (i.e., α2–3- versus α2–6-linked sialyllactosides) of the neuraminidase. Herein we report a glycoarray-based assay and a MALDI-TOF study for assessing the activity and specificity of two influenza neuraminidases on whole viruses. The human A(H3N2) and avian A(H5N2) neuraminidase activities were investigated. The results from both approaches demonstrated that α2–3 sialyllactoside was a better substrate than α2–6 sialyllactoside for both viruses and that H5N2 virus had a lower hydrolytic activity than H3N2.

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