Analysis of interaction between M2-2 protein of human respiratory syncytial virus with quercetin and murin

Abstract

The Human Respiratory Syncytial Virus (RSV) is the major causative agent of acute respiratory infections (ARI), such as pneumonia and bronchiolitis. It is a virus of the Paramyxoviridae family, with helical symmetry and lipid envelope. It has a single-stranded RNA genome, not segmented, with 10 coding genes. One of the factors that contribute to success in viral replication is the interaction of M2-2 protein with ribosomal complex leading to phase change of viral activity and may inhibit or induce the replication of the virus according to the concentration of mRNA. Such protein can act by controlling the mRNA synthesis and thus effectively participate in the viral replication process and to help shape the work in the silencing of the ribonucleoprotein complex (RNP) of RSV. Thus, the binder to perform an interaction with the M2-2 is a potential candidate to prevent the M2-2 interactions with RNA and other proteins, thereby preventing the success of viral replication. To investigate the interaction of a potential inhibitor of the M2-2 activity the objective of the project is to clone, express, and purify the M2-2 protein hRSV and then determine its physicochemical characteristic of interaction with Quercetin and Morina respectively, under different temperature conditions. For cloning and expression of the M2-2 gene have been chosen vector pET28a and Escherichia coli BL21 strain. The expressed protein will be purified by affinity chromatography and ion-exchange chromatography. The protein interaction measurements with binders (Quercetin and Morina) will be carried out by fluorescence spectroscopy at different temperatures to characterize the nature of the interaction. The compositi¬¬on of secondary structure and thermal stability will be determined through the technique of circular dichroism spectroscopy. Thus, the results obtained in this project will make it possible to propose a therapeutic target for combat viral infection process hRSV and still will point out a potential inhibitor of interaction for M2-2 and consequently leading to blocking viral replication.