title: Anomeric specificity and protein-substrate interactions support the 3D model for the hemagglutinin-neuraminidase from sendai virus.
abstract: The 3D structure of paramyxovirus hemagglutinin-neuraminidase has not yet been resolved; however, a theoretical model has been built by using influenza virus and bacterial neuraminidases as template [V. C. Epa (1997) Proteins Struct. Funct. Gen. 29, 264-281]. Two common features of the catalytic mechanism of the neuraminidases of known 3D structure are the anomeric specificity and the involvement of a tyrosine residue in the stabilization of the transition state. These key features have been investigated on the water-soluble ectodomain of the hemagglutinin-neuraminidase from Sendai virus (cHN). The anomeric specificity of the hydrolysis of the substrate by cHN has been investigated by NMR spectroscopy. The immediate product of the reaction was the alpha-anomer, meaning that cHN belongs between glycohydrolases retaining anomeric configuration like influenza virus neuraminidase. Measurements of the UV difference spectrum upon binding of the substrate analogue 2,3-dehydro 2-deossi N-acetyl neuraminic acid indicate the ionization of a tyrosine residue and decreased polarity in the environment of a tryptophan residue. Functional significance of the spectral data was derived from the known structure of influenza neuraminidase, where a tyrosinate ion is involved in the stabilization of the transition-state carbonium ion, and a tryptophan residue is involved in the binding of the acetyl moiety of the substrate. The data give experimental support to the 3D model of paramyxovirus neuraminidase.
authors.affiliation: Dipartimento di Biochimica e Biologia Molecolare, Università di Ferrara, via Borsari 46, Ferrara, 44100, Italy.
authors.forename: T
authors.initials: T
authors.lastname: Bellini
authors.forename: C
authors.initials: C
authors.lastname: Pasti
authors.forename: M C
authors.initials: MC
authors.lastname: Manfrinato
authors.forename: M
authors.initials: M
authors.lastname: Tomasi
authors.forename: F
authors.initials: F
authors.lastname: Dallocchio
mesh_terms: D020134:Catalytic Domain; D015707:HN Protein / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D007536:Isomerism; D008958:Models, Molecular; D009438:Neuraminic Acids / Q000378:metabolism; D009439:Neuraminidase / Q000737:chemistry* / Q000378:metabolism; D019906:Nuclear Magnetic Resonance, Biomolecular; D018108:Respirovirus / Q000201:enzymology*; D013379:Substrate Specificity
chemical_list: D015707:HN Protein; D009438:Neuraminic Acids; C045864:2,3-dehydroneuraminic acid; D009439:Neuraminidase