Expression, purification and characterization of Macrobrachium rosenbergii nodavirus capsid protein

This is the research work ( from Ms. Li Chuin Chong ( She completed her first degree (BSc) in Biomedical Sciences from University Putra Malaysia (UPM) and then pursued a master's degree (MSc) in Bioinformatics at Perdana University School of Data Sciences (PU-ScDS), Malaysia. She is very enthusiastic and passionate about science. 

Short summary of the respective article

Macrobrachium rosenbergii nodavirus (MrNV) is the causative agent of white tail disease (WTD) which seriously impedes the production of the giant freshwater prawn and has a major economic impact. MrNV contains two segmented RNA molecules, which encode the RNA dependent RNA polymerase (RdRp) and the capsid protein (MrNV-CP) containing 371 amino acid residues. MrNV-CP comprises of the Shell (S) and the Protruding (P) domains, ranging from amino acid residues 1–252 and 253–371, respectively. The P-domain assembles into dimeric protruding spikes, and it is believed to be involved in host cell attachment and internalization. In this study, the recombinant P-domain of MrNV-CP was successfully cloned and expressed in Escherichia coli, purified with an immobilized metal affinity chromatography (IMAC) and size exclusion chromatography (SEC) up to ~90% purity. Characterization of the purified recombinant P-domain with SEC revealed that it formed dimers, and dynamic light scattering (DLS) analysis demonstrated that the hydrodynamic diameter of the dimers was ~6 nm. Circular dichroism (CD) analysis showed that the P-domain contained 67.9% of beta-sheets, but without alpha-helical structures. This is in good agreement with the cryo-electron microscopic analysis of MrNV which demonstrated that the P-domain contains only beta-stranded structures. This research findings of this study provide essential information for the production of the P-domain of MrNV-CP that will aid future studies particularly studies that will shed light on anti-viral drug discovery and provide an understanding of virus-host interactions and the viral pathogenicity.

The P-domain of MrNV-CP was expressed as a soluble protein in E. coli. The recombinant Pdomain formed dimers with a hydrodynamic diameter ~6 nm which resemble the full-length capsid protein. This research data provide crucial biophysical parameters for downstream analyses of the P-domain of MrNV-CP such as structural analysis using X-ray crystallography or nuclear magnetic resonance. Availability of a high-resolution 3D structure of the P-domain is essential to elucidate the mechanisms involved in host cell recognition and penetration.

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