Biophysical characterization of the interaction between M2-1 protein of hRSV and quercetin

hRSV is the major causative agent of acute respiratory infections. Among its eleven proteins, M2-1 is a transcription antiterminator, making it an interesting target for antivirals. Quercetin is a flavonol which inhibits some virus infectivity and replication. In the present work, the M2-1 gene was...

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Detalles Bibliográficos
Autores: Teixeira, Thiago Salem Pançonato [UNESP], Caruso, Ícaro Putinhon [UNESP], Lopes, Bruno Rafael Pereira [UNESP], Regasini, Luis Octávio [UNESP], Toledo, Karina Alves de [UNESP], Fossey, Marcelo Andrés [UNESP], Souza, Fátima Pereira de [UNESP]
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2017
País:Brasil
Institución:Universidade Estadual Paulista (UNESP)
Repositorio:Repositório Institucional da UNESP
Idioma:inglés
OAI Identifier:oai:repositorio.unesp.br:11449/173810
Acceso en línea:http://dx.doi.org/10.1016/j.ijbiomac.2016.11.033
http://hdl.handle.net/11449/173810
Access Level:acceso abierto
Palabra clave:Biophysical analyses
hRSV
M2-1
Molecular docking
Quercetin
Descripción
Sumario:hRSV is the major causative agent of acute respiratory infections. Among its eleven proteins, M2-1 is a transcription antiterminator, making it an interesting target for antivirals. Quercetin is a flavonol which inhibits some virus infectivity and replication. In the present work, the M2-1 gene was cloned, expressed and the protein was purified. Thermal stability and secondary structure were analyzed by circular dichroism and the interaction with Quercetin was evaluated by fluorescence spectroscopy. Molecular docking experiments were performed to understand this mechanism of interaction. The purified protein is mainly composed of α-helix, with a melting temperature of 328.6 K (≈55 °C). M2-1 titration with Quercetin showed it interacts with two sites, one with a strong constant association K1 (site 1 ≈ 1.5 × 106 M−1) by electrostatic interactions, and another with a weak constant association K2 (site 2 ≈ 1.1 × 105 M−1) by a hydrophobic interaction. Ligand's docking shows it interacts with the N-terminus face in a more polar pocket and, between the domains of oligomerization and RNA and P protein interaction, in a more hydrophobic pocket, as predicted by experimental data. Therefore, we postulated this ligand could be interacting with important domains of the protein, avoiding viral replication and budding.