The Correlation between the Substitution of Superficial Residues on Subviral Particles of the Infectious Bursal Disease Virus and their Interactions with Nickel Ions

• Main Authors: Ren-Hao Yu, 余仁豪
• Other Authors: Min-Ying Wang
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/75814054842962960578

碩士 === 國立中興大學 === 生物科技學研究所 === 98 === VP2 is the major viral capsid protein of infectious bursal disease virus (IBDV), a causative agent of a highly contagious immunosuppressive disease in young chickens. In vivo expression of VP2 can cause the formation of icosahedral (T=1) subviral particles (SVPs). Monomeric VP2 protein contains three domains, ie., protrusion (P), shell(S) and base (B) domain. Outward P domain (aa.202~341) is to compose the loop and β-sheet. In this study, to simulate the protein structure of iran strain IR01 VP2 base on local strain P3009 VP2 by using homology modeling, then employed Surface Racer 5.0 and Discovery Studio 2.5 to calculate the exposure area and percent solvent accessibility of side chains of P3009 VP2 and IR01 VP2, respectively. Previous results demonstrated that SVPs can bind to Ni ion by His253 on the DE loop of VP2 P domain of SVP, the specific amino acid were substituted with histidine via site-directed mutagenesis to generate few variants. P3009 VP2, IR01 VP2 mutants were separated by sucrose gradient centrifugation. In the results, themutants have same distribution campare with the WT SVP, the follow-up transmission electron microscopy identified P3009 and IR01 VP2 particles structures have similar diameter with wild type SVP . To test the adsorption of each mutants by immobilized metal affinity chromatography, the results showed the D279H, Q324H on the β-sheet structure of P domain of P3009 VP2 provide the adsorptive to Ni ion and S251H, G285H and Q324H of IR01 VP2 were carried out with the nickel ion adsorption, respectively. Conclusion of this study, the D279H, Q324H in the β-sheet structure of P3009 VP2 provide the adsorption with IMAC and adsorption results of IR01 VP2 showed that credibility of IR01 subvirus particle model from homology modeling.