Tropical Journal of Pharmaceutical Research

Original Research Article | OPEN ACCESS

Identification of putative vero cell protein(s) that bind specifically to recombinant envelope protein of dengue virus type 2

Seema Zargar¹ , Tanveer A Wani², SK Jain³

¹Department of Biochemistry, King Saud University, PO Box 22452; ²Department of Pharmaceutical Chemistry, College of Pharmacy, King Saud University, Riyadh, 11451, Kingdom of Saudi Arabia; ³Department of Biotechnology, Faculty of Science, Jamia Hamdard, New Delhi, India.

For correspondence:- Seema Zargar Email: szargar@ksu.edu.sa Tel:+966118051692

Received: 27 December 2014 Accepted: 30 April 2015 Published: 29 June 2015

Citation: Zargar S, Wani TA, Jain S. Identification of putative vero cell protein(s) that bind specifically to recombinant envelope protein of dengue virus type 2. Trop J Pharm Res 2015; 14(6):997-1003 doi: 10.4314/tjpr.v14i6.9

© 2015 The authors.
This is an Open Access article that uses a funding model which does not charge readers or their institutions for access and distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0) and the Budapest Open Access Initiative (http://www.budapestopenaccessinitiative.org/read), which permit unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited..

Abstract

Purpose: To identify protein targets in host (vero) cell since there is currently no therapy or a licensed tetravalent vaccine to combat all the four virus serotypes of dengue virus.

Methods: The domain III of the dengue virus encoded envelope protein was expressed in pET28a expression vector and the purified recombinant protein was labeled with biotin without altering its immunogenicity. Vero cell proteins on nitrocellulose membrane reacted with recombinant envelope protein domain III to identify viral target proteins in vero cells.

Results: The 45 KDa, 43 KDa and 30 KDa plasma membrane proteins were identified as viral envelope targets. Competitive binding assay showed these proteins competing with dengue virus binding. MTT assay indicate that viability of vero cells increases in cultures pretreated with 45 KDa, 43 KDa and 30 KDa proteins before dengue infection.

Conclusion: These results indicate the possible role of these proteins in viral binding to vero cells. The study provides a preliminary insight that would aid in determining the target epitopes against protein E domain III of dengue virus and hence, formulation of a vaccine for preparing neutralizing antibodies.

Keywords: Dengue virus envelope, Biotinylation, Ni-NTA purification, Target epitopes, Plaque assay, Competitive blocking assay