Recombinant turnip yellow mosaic virus coat protein as a potential nanocarrier.

Autor: Tan FH; School of Biosciences, Faculty of Health and Medical Sciences, Taylor's University, Subang Jaya, Selangor, Malaysia., Kong JC; School of Biosciences, Faculty of Health and Medical Sciences, Taylor's University, Subang Jaya, Selangor, Malaysia., Ng JF; School of Biosciences, Faculty of Health and Medical Sciences, Taylor's University, Subang Jaya, Selangor, Malaysia., Alitheen NB; Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia., Wong CL; Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia., Yong CY; Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia., Lee KW; School of Biosciences, Faculty of Health and Medical Sciences, Taylor's University, Subang Jaya, Selangor, Malaysia.
Jazyk: angličtina
Zdroj: Journal of applied microbiology [J Appl Microbiol] 2021 Oct; Vol. 131 (4), pp. 2072-2080. Date of Electronic Publication: 2021 Mar 06.
DOI: 10.1111/jam.15048
Abstrakt: Aims: To display a short peptide (GSRSHHHHHH) at the C-terminal end of turnip yellow mosaic virus coat protein (TYMVc) and to study its assembly into virus-like particles (TYMVcHis 6 VLPs).
Methods and Results: In this study, recombinant TYMVcHis 6 expressed in Escherichia coli self-assembled into VLPs of approximately 30-32 nm. SDS-PAGE and Western blot analysis of protein fractions from the immobilized metal affinity chromatography (IMAC) showed that TYMVcHis 6 VLPs interacted strongly with nickel ligands in IMAC column, suggesting that the fusion peptide is protruding out from the surface of VLPs. These VLPs are highly stable over a wide pH range from 3·0 to 11·0 at different temperatures. At pH 11·0, specifically, the VLPs remained intact up to 75°C. Additionally, the disassembly and reassembly of TYMVcHis 6 VLPs were studied in vitro. Dynamic light scattering and transmission electron microscopy analysis revealed that TYMVcHis 6 VLPs were dissociated by 7 mol l -1 urea and 2 mol l -1 guanidine hydrochloride (GdnHCl) without impairing their reassembly property.
Conclusions: A 10-residue peptide was successfully displayed on the surface of TYMVcHis 6 VLPs. This chimera demonstrated high stability under extreme thermal conditions with varying pH and was able to dissociate and reassociate into VLPs by chemical denaturants.
Significance and Impact of the Study: This is the first C-terminally modified TYMVc produced in E.  coli. The C-terminal tail which is exposed on the surface can be exploited as a useful site to display multiple copies of functional ligands. The ability of the chimeric VLPs to self-assemble after undergo chemical denaturation indicates its potential role to serve as a nanocarrier for use in targeted drug delivery.
(© 2021 The Society for Applied Microbiology.)
Databáze: MEDLINE
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