Cloning, expression and analysis of the interaction of the M2-1 protein of HRSV with Flavonides: investigation for targets in blocking viral replication

• Grant number: 13/24355-2
• Support Opportunities: Regular Research Grants
• Start date: August 01, 2014
• End date: July 31, 2016
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Fátima Pereira de Souza
• Grantee: Fátima Pereira de Souza
• Host Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil
• Associated researchers: Fernando Alves de Melo ; Karina Alves de Toledo ; Luis Octávio Regasini ; Marcelo Andrés Fossey

Abstract

Human Respiratory Syncytial Virus (hRSV) is the major causative agent of acute respiratory infections (ARIs) such as pneumonia and bronchiolitis. This is a virus of the Paramyxoviridae family with helical symmetry and lipid envelope. It has a single strand RNA genome, not segmented, with 10 coding genes. One of the factors that contribute to success in viral replication is the interaction M2-1 protein with P, M protein and RNA viral. This protein may act inhibiting or neutralizing various steps of the pathway of interferons or to assist in silencing ribonucleoprotein complex (RNP) of RSV. Thus a linkage that render interaction with M2-1 is a potential candidate to prevent interaction of M2-1 with other proteins, thereby preventing the success of viral replication. The aim of the research is to clone, express and purify the M2-1 protein of hRSV and then determine physicochemical characteristics of the interaction with flavonoids in temperature different to investigate the interaction of a potential candidate for inhibiting the activity of the M2-1. The M2-1 gene will be cloned in vector pET28A and expressed in Escherichia coli strain BL21. The expressed protein will be purified on affinity chromatography, thermal stability and interaction with flavonoids will be analyzed by spectroscopy. The results of this study will be unveil protein stability conditions and the interaction site of small ligands, allowing to obtain interaction model, which could lead to potent therapeutic models. As a long term scope, the protein structure determination will be used for refined experiments that will provide better understanding of the flavonoids binding sites. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

GUIMARAES, GIOVANA C.; PIVA, HEMILY R. M.; ARAUJO, GABRIELA C.; LIMA, CAROLINE S.; REGASINI, LUIS O.; DE MELO, FERNANDO A.; FOSSEY, MARCELO A.; CARUSO, ICARO P.; SOUZA, FATIMA P.. Binding investigation between M2-1protein from hRSV and acetylated quercetin derivatives: H-1 NMR, fluorescence spectroscopy, and molecular docking. International Journal of Biological Macromolecules, v. 111, p. 33-38, MAY 2018. (13/24355-2, 09/53989-4)

PANCONATO TEIXEIRA, THIAGO SALEM; CARUSO, ICARO PUTINHON; PEREIRA LOPES, BRUNO RAFAEL; REGASINI, LUIS OCTAVIO; DE TOLEDO, KARINA ALVES; FOSSEY, MARCELO ANDRES; DE SOUZA, FATIMA PEREIRA. Biophysical characterization of the interaction between M2-1 protein of hRSV and quercetin. International Journal of Biological Macromolecules, v. 95, p. 63-71, FEB 2017. (13/24355-2)

DE LOURENCO, ISABELLA OTTENIO; QUINTINO, EVELYN TOSCANO PEDROSO; PEREIRA, MATHEUS HENRIQUE; LIMA, CAROLINE SPRENGEL; ARAUJO, GABRIELA CAMPOS; REGASINI, LUIS OCTAVIO; MELO, FERNANDO ALVES DE; SOUZA, FATIMA PEREIRA DE; FOSSEY, MARCELO ANDRES; CARUSO, ICARO PUTINHON. Biophysical studies of the interaction of hRSV Non-Structural 1 protein with natural flavonoids and their acetylated derivatives by spectroscopic techniques and computational simulations. SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY, v. 283, p. 12-pg., 2022-08-13. (13/24355-2, 10/18169-3, 15/18868-2)