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Use of a lambda gt11 expression library to localize a neutralizing antibody-binding site in glycoprotein E2 of Sindbis virus

Wang, Kang-Sheng and Strauss, James H. (1991) Use of a lambda gt11 expression library to localize a neutralizing antibody-binding site in glycoprotein E2 of Sindbis virus. Journal of Virology, 65 (12). pp. 7037-7040. ISSN 0022-538X. http://resolver.caltech.edu/CaltechAUTHORS:WANjvir91

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Abstract

The Sindbis virus envelope contains two species of integral membrane glycoproteins, E1 and E2. These proteins form heterodimers, and three dimeric units assemble to form spikes incorporated into the viral surface which play an important role in the specific attachment of Sindbis virus to host cells. To map the neutralization epitopes on the surface of the virus, we constructed a lambda gt11 expression library with cDNA inserts 100 to 300 nucleotides long obtained from randomly primed synthesis on Sindbis virus genomic RNA. This library was screened with five different neutralizing monoclonal antibodies (MAbs) specific for E2 (MAbs 50, 51, 49, 18, and 23) and with one neutralizing MAb specific for E1 (MAb 33). When 10(6) lambda gt11 plaques were screened with each antibody, four positive clones that reacted with E2-specific MAb 23 were found. These four clones contained overlapping inserts from glycoprotein E2; the domain from residues 173 to 220 of glycoprotein E2 was present in all inserts, and we concluded that this region contains the neutralization epitope recognized by the antibody. No clones that reacted with the other antibodies examined were found, and we concluded that these antibodies probably recognize conformational epitopes not present in the lambda gt11 library. We suggest that the E2 domain from residues 173 to 220 is a major antigenic determinant of Sindbis virus and that this domain is important for virus attachment to cells.


Item Type:Article
Additional Information:Copyright © 1991 by the American Society for Microbiology. Received 14 June 1991/Accepted 28 August 1991 We are grateful to R. Kuhn, A. Schmaljohn, R. Weir, and E. Strauss for help and advice during the conduct of these experiments and preparation of the manuscript. This work was supported by grant AI 10793 from the NIH and by contract DAMD17-90-C-0050 from the U.S. Army Medical Research and Development Command.
Record Number:CaltechAUTHORS:WANjvir91
Persistent URL:http://resolver.caltech.edu/CaltechAUTHORS:WANjvir91
Alternative URL:http://jvi.asm.org/cgi/content/abstract/65/12/7037
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:5989
Collection:CaltechAUTHORS
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Deposited On:10 Nov 2006
Last Modified:26 Dec 2012 09:16

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