Abstract

Characterization of Glycoprotein E C-End of West Nile Virus and Evaluation of Its Interaction Force with αVβ3 Integrin as Putative Cellular Receptor

M. V. Bogachek¹, B. N. Zaitsev¹, S. K. Sekatskii², E. V. Protopopova¹, V. A. Ternovoi¹, A. V. Ivanova¹, A. V. Kachko¹, V. A. Ivanisenko³, G. Dietler², and V. B. Loktev^1*

¹FSRI State Research Center of Virology and Biotechnology “Vector”, 630559 Koltsovo, Novosibirsk Region, Russia; fax: (383) 336-7409; E-mail: loktev@vector.nsc.ru

²Laboratoire de Physique de la Matiere Vivante, IPSB, BSP, Ecole Polytechnique Federale de Lausanne, CH 1015, Lausanne, Switzerland; fax: (4121) 693-0422; E-mail: Serguei.Sekatski@epfl.ch

³Institute of Cytology and Genetics, Siberian Branch of the Russian Academy of Sciences, ul. Lavrentieva 10, 630090 Novosibirsk, Russia; fax: (383) 333-1278; E-mail: salix@bionet.nsc.ru

^* To whom correspondence should be addressed.

Received May 25, 2009; Revision received September 13, 2009
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–αVβ3 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 αVβ3 integrin) during virus entry.
KEY WORDS: West Nile virus, glycoprotein E, atomic force spectroscopy, αVβ3 integrin
DOI: 10.1134/S0006297910040115

Characterization of Glycoprotein E C-End of West Nile Virus and Evaluation of Its Interaction Force with αVβ3 Integrin as Putative Cellular Receptor

M. V. Bogachek1, B. N. Zaitsev1, S. K. Sekatskii2, E. V. Protopopova1, V. A. Ternovoi1, A. V. Ivanova1, A. V. Kachko1, V. A. Ivanisenko3, G. Dietler2, and V. B. Loktev1*

M. V. Bogachek¹, B. N. Zaitsev¹, S. K. Sekatskii², E. V. Protopopova¹, V. A. Ternovoi¹, A. V. Ivanova¹, A. V. Kachko¹, V. A. Ivanisenko³, G. Dietler², and V. B. Loktev^1*