title: Molecular modeling and conformational analysis of native and refolded viral genome-linked protein of cardamom mosaic virus.
abstract: The viral genome-linked protein (VPg) of Potyviruses is covalently attached to the 5' end of the genomic RNA. Towards biophysical characterization, the VPg coding region of Cardamom mosaic virus (CdMV) was amplified from the cDNA and expressed in E. coli. Most of the expressed VPg aggregated as inclusion bodies that were solubilized with urea and refolded with L-arginine hydrochloride. The various forms of CdMV VPg (native, denatured and refolded) were purified and the conformational variations between these forms were observed with fluorescence spectroscopy. Native and refolded CdMV VPg showed unordered secondary structure in the circular dichroism (CD) spectrum. The model of CdMV VPg was built based on the crystal structure of phosphotriesterase (from Pseudomonas diminuta), which had the maximum sequence homology with VPg to identify the arrangement of conserved amino acids in the protein to study the functional diversity of VPg. This is the first report on the VPg of CdMV, which is classified as a new member of the Macluravirus genus of the Potyviridae family.
authors.affiliation: Department of Plant Biotechnology, School of Biotechnology, Madurai Kamaraj University, Madurai 625021, Tamil Nadu, India.
authors.forename: T
authors.initials: T
authors.lastname: Jebasingh
authors.forename: M
authors.initials: M
authors.lastname: Jose
authors.forename: A Kasin
authors.initials: AK
authors.lastname: Yadunandam
authors.forename: S
authors.initials: S
authors.lastname: Backiyarani
authors.forename: K V
authors.initials: KV
authors.lastname: Srividhya
authors.forename: S
authors.initials: S
authors.lastname: Krishnaswamy
authors.forename: R
authors.initials: R
authors.lastname: Usha
mesh_terms: D002942:Circular Dichroism; D028523:Elettaria / Q000378:metabolism / Q000821:virology*; D016679:Genome, Viral / Q000235:genetics*; D002479:Inclusion Bodies / Q000235:genetics* / Q000378:metabolism / Q000821:virology*; D008958:Models, Molecular; D009029:Mosaic Viruses / Q000235:genetics / Q000378:metabolism; D010942:Plant Viruses / Q000235:genetics / Q000378:metabolism; D017800:Potyvirus / Q000235:genetics / Q000378:metabolism; D058849:Protein Refolding; D017433:Protein Structure, Secondary; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D016601:RNA-Binding Proteins; D014764:Viral Proteins