title: Quantifying the specific binding between West Nile virus envelope domain III protein and the cellular receptor alphaVbeta3 integrin.
abstract: A previous study has illustrated that the alphaVbeta3 integrin served as the functional receptor for West Nile virus (WNV) entry into cells. Domain III (DIII) of WNV envelope protein (E) was postulated to mediate virus binding to the cellular receptor. In this study, the specificity and affinity binding of WNV E DIII protein to alphaVbeta3 integrin was confirmed with co-immunoprecipitation and receptor competition assay. Binding of WNV E DIII protein to alphaVbeta3 integrin induced the phosphorylation of focal adhesion kinase that is required to mediate ligand-receptor internalization into cells. A novel platform was then developed using the atomic force microscopy to measure this specific binding force between WNV E DIII protein and the cellular receptor, alphaVbeta3 integrin. The single protein pair-interacting force measured was in the range of 45 +/- 5 piconewtons. This interacting force was highly specific as minimal force was measured in the WNV E DIII protein interaction with alphaVbeta5 integrin molecules and heparan sulfate. These experiments provided an insight to quantitate virus-receptor interaction. Force measurement using atomic force microscopy can serve to quantitatively analyze the effect of candidate drugs that modulate virus-host receptor affinity.
authors.affiliation: Flavivirology Laboratory, Department of Microbiology, National University of Singapore, Singapore 117597.
authors.forename: Jason Wei-Ming
authors.initials: JW
authors.lastname: Lee
authors.forename: Justin Jang-Hann
authors.initials: JJ
authors.lastname: Chu
authors.forename: Mah-Lee
authors.initials: ML
authors.lastname: Ng
mesh_terms: D045744:Cell Line, Tumor; D015686:Gene Products, env / Q000378:metabolism*; D006801:Humans; D039302:Integrin alphaVbeta3 / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008545:Melanoma; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D014902:West Nile virus / Q000378:metabolism*
chemical_list: D015686:Gene Products, env; D039302:Integrin alphaVbeta3