Preprint Article Version 1 This version is not peer-reviewed

Characterization of Recombinant Poliovirus 2A Protease; A Potential Anti-Viral Drug Target

Version 1 : Received: 21 November 2018 / Approved: 22 November 2018 / Online: 22 November 2018 (14:41:00 CET)

How to cite: Younus, A.; Bhatti, M.F.; Virk, N.; Arshad, M.; Janjua, H.; Coutts, R. Characterization of Recombinant Poliovirus 2A Protease; A Potential Anti-Viral Drug Target. Preprints 2018, 2018110550 (doi: 10.20944/preprints201811.0550.v1). Younus, A.; Bhatti, M.F.; Virk, N.; Arshad, M.; Janjua, H.; Coutts, R. Characterization of Recombinant Poliovirus 2A Protease; A Potential Anti-Viral Drug Target. Preprints 2018, 2018110550 (doi: 10.20944/preprints201811.0550.v1).

Abstract

Poliovirus 2A protease (PV2Apro) plays a vital role in viral replication and down-regulation of host cell protein synthesis. In order to understand more concerning PV2Apro, the protein was over-expressed in bacteria following amplification using sense and antisense primers and cloning in pET15b. Several expression hosts were tested and BL21 (DE3) pLysS cells gave the best expression of PV2Apro with minimal unwanted protein expression following IPTG induction. The 2Apro protein was purified to homogeneity using column chromatography, its solubility determined and its molecular weight and composition determined by MALDI-TOF mass spectrometry. The protease was found in the insoluble fraction and the purified protein had a slightly lower molecular weight than predicted. Moreover, three dimensional structure was modelled using template 1z8r with 58% identity and validated using ramachandran plot. Results revealed that most of the residues lie in favoured and allowed regions. These findings could help in a better understanding of PV2Apro structure and inhibition thus, highlighting potential targets for antiviral drug development.

Subject Areas

poliovirus 2A protease (PV2Apro); expression; purification; MALDI-TOF; 3D structure

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