title: Characterization of glycoprotein E C-end of West Nile virus and evaluation of its interaction force with alphaVbeta3 integrin as putative cellular receptor.
abstract: Recombinant polypeptide containing the 260-466 amino acid sequence of West Nile virus (WNV) strain LEIV-Vlg99-27889-human glycoprotein E (gpE, E(260-466)) was constructed. Immunochemical similarity between the E(260-466) and gpE of WNV was proven by enzyme immunoassay (EIA), immunoblot, competitive EIA, hemagglutination inhibition, and neutralization tests using polyclonal and monoclonal antibodies against the viral gpE and recombinant E(260-466). Polypeptide E(260-466) induced formation of virus neutralizing and cross-reactive antibodies that were interactive with various epitopes of this recombinant protein. It is shown by evaluation of the interaction of E(260-466) with one of the proposed cell receptors of WNV that average E(260-466)-alphaVbeta3 integrin-specific interaction force measured using atomic force spectroscopy was 80 and 140 pN for single and double interactions, correspondingly. Taken together with previously described interaction between laminin-binding protein (LBP) and WNV gpE domain II, it is proposed that WNV gpE can interact specifically with two cellular proteins (LBP and alphaVbeta3 integrin) during virus entry.
authors.affiliation: FSRI State Research Center of Virology and Biotechnology Vector, Koltsovo, Novosibirsk Region, 630559, Russia.
authors.forename: M V
authors.initials: MV
authors.lastname: Bogachek
authors.forename: B N
authors.initials: BN
authors.lastname: Zaitsev
authors.forename: S K
authors.initials: SK
authors.lastname: Sekatskii
authors.forename: E V
authors.initials: EV
authors.lastname: Protopopova
authors.forename: V A
authors.initials: VA
authors.lastname: Ternovoi
authors.forename: A V
authors.initials: AV
authors.lastname: Ivanova
authors.forename: A V
authors.initials: AV
authors.lastname: Kachko
authors.forename: V A
authors.initials: VA
authors.lastname: Ivanisenko
authors.forename: G
authors.initials: G
authors.lastname: Dietler
authors.forename: V B
authors.initials: VB
authors.lastname: Loktev
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000276:immunology; D000939:Epitopes / Q000737:chemistry / Q000276:immunology; D006801:Humans; D039482:Integrin alpha5 / Q000737:chemistry* / Q000378:metabolism; D018625:Microscopy, Atomic Force; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014759:Viral Envelope Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D014902:West Nile virus / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D000939:Epitopes; D039482:Integrin alpha5; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins