147 articles – 175 references  [version française]
Detailed view Article in peer-reviewed journal
ChemBioChem 12, 13 (2011) 2071-80
Measurement of enzymatic activity and specificity of human and avian influenza neuraminidases from whole virus by glycoarray and MALDI-TOF mass spectrometry.
Gwladys Pourceau1, Yann Chevolot2, Alice Goudot2, Fabienne Giroux3, Albert Meyer1, Vincent Moulés3, Bruno Lina3, Samy Cecioni4, Sébastien Vidal4, Hai Yu5, Xi Chen5, Olivier Ferraris3, Jean-Pierre Praly4, Eliane Souteyrand2, Jean-Jacques Vasseur1, François Morvan1

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.
1:  IBMM - Institut des Biomolécules Max Mousseron
2:  INL - Institut des nanotechnologies de Lyon - Site d'Ecully
3:  VirPath - Virologie et Pathologie Humaine
4:  ICBMS - Institut de Chimie et Biochimie Moléculaires et Supramoléculaires
5:  University of California