title: Measurement of enzymatic activity and specificity of human and avian influenza neuraminidases from whole virus by glycoarray and MALDI-TOF mass spectrometry.
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.
authors: IBMM UMR 5247 CNRS UM1 UM2, Université de Montpellier 2, Place Eugène Bataillon, 34095 Montpellier Cedex 5, France.
authors: Gwladys
authors: G
authors: Pourceau
authors: Yann
authors: Y
authors: Chevolot
authors: Alice
authors: A
authors: Goudot
authors: Fabienne
authors: F
authors: Giroux
authors: Albert
authors: A
authors: Meyer
authors: Vincent
authors: V
authors: Moulés
authors: Bruno
authors: B
authors: Lina
authors: Samy
authors: S
authors: Cecioni
authors: Sébastien
authors: S
authors: Vidal
authors: Hai
authors: H
authors: Yu
authors: Xi
authors: X
authors: Chen
authors: Olivier
authors: O
authors: Ferraris
authors: Jean-Pierre
authors: JP
authors: Praly
authors: Eliane
authors: E
authors: Souteyrand
authors: Jean-Jacques
authors: JJ
authors: Vasseur
authors: François
authors: F
authors: Morvan
mesh_terms: D000818:Animals; D001717:Birds; D006801:Humans; D053122:Influenza A Virus, H3N2 Subtype / Q000201:enzymology*; D053125:Influenza A Virus, H5N2 Subtype / Q000201:enzymology*; D005585:Influenza in Birds / Q000821:virology; D007251:Influenza, Human / Q000821:virology; D046228:Microarray Analysis / Q000379:methods; D009439:Neuraminidase / Q000378:metabolism*; D013045:Species Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D009439:Neuraminidase