title: Biophysical characterization of G protein ectodomain of group B human respiratory syncytial virus from E. coli.
abstract: Human respiratory syncytial virus (hRSV) is an important pathogen of acute respiratory tract infection. The G protein of hRSV is a transmembrane glycoprotein that is a neutralizing antigen and is thus a vaccine candidate. In this study, synthetic codon optimized ectodomain G protein [G(ΔTM)] of BA genotype of group B hRSV was cloned, expressed, and characterized using biophysical techniques. The molar absorption coefficient and mean residue ellipticity at 222 nm ([θ]222) of G (ΔTM) was found to be 7950 M(-1) cm(-1) and -19701.7 deg cm(2) dmol(-1) respectively. It was concluded that G(ΔTM) mainly consist of α-helix (74.9%) with some amount of β-sheet (4%). The protein was stable up to 85°C without any transition curve. However, heat-induced denaturation of G(ΔTM) resulted in total loss of β-sheet whereas not much change was observed in the α-helix part of the secondary structure. It was concluded that G(ΔTM) is an α-helical protein and it is highly stable at high temperature, but could be easily denatured using high concentrations of GdmCl/urea or acidic condition. This is the first investigation of cloning, expression, and characterization of G(ΔTM) of BA viruses from India. Structural characterization of G protein will assist in drug designing and vaccine development for hRSV.
authors.affiliation: a Centre for Interdisciplinary Research in Basic Sciences , Jamia Millia Islamia , New Delhi , India.
authors.forename: Wajihul Hasan
authors.initials: WH
authors.lastname: Khan
authors.affiliation: b Department of Microbiology , All India Institute of Medical Sciences , New Delhi , India.
authors.forename: V L N Raghuram
authors.initials: VL
authors.lastname: Srungaram
authors.affiliation: a Centre for Interdisciplinary Research in Basic Sciences , Jamia Millia Islamia , New Delhi , India.
authors.forename: Asimul
authors.initials: A
authors.lastname: Islam
authors.affiliation: a Centre for Interdisciplinary Research in Basic Sciences , Jamia Millia Islamia , New Delhi , India.
authors.forename: Ilyas
authors.initials: I
authors.lastname: Beg
authors.affiliation: a Centre for Interdisciplinary Research in Basic Sciences , Jamia Millia Islamia , New Delhi , India.
authors.forename: Md Shakir H
authors.initials: MS
authors.lastname: Haider
authors.affiliation: a Centre for Interdisciplinary Research in Basic Sciences , Jamia Millia Islamia , New Delhi , India.
authors.forename: Faizan
authors.initials: F
authors.lastname: Ahmad
authors.affiliation: b Department of Microbiology , All India Institute of Medical Sciences , New Delhi , India.
authors.forename: Shobha
authors.initials: S
authors.lastname: Broor
authors.affiliation: a Centre for Interdisciplinary Research in Basic Sciences , Jamia Millia Islamia , New Delhi , India.
authors.forename: Shama
authors.initials: S
authors.lastname: Parveen
mesh_terms: D000595:Amino Acid Sequence; D001703:Biophysics; D004926:Escherichia coli / Q000235:genetics*; D019204:GTP-Binding Proteins / Q000502:physiology*; D006801:Humans; D012136:Respiratory Syncytial Viruses / Q000235:genetics / Q000472:pathogenicity*; D017386:Sequence Homology, Amino Acid
chemical_list: D019204:GTP-Binding Proteins
keywords: BA genotype; biophysical characterization; ectodomain G protein; human respiratory syncytial virus