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Journal of Virology, June 2002, p. 5315-5325, Vol. 76, No. 11
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.11.5315-5325.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Envelope Glycoprotein Incorporation, Not Shedding of Surface Envelope Glycoprotein (gp120/SU), Is the Primary Determinant of SU Content of Purified Human Immunodeficiency Virus Type 1 and Simian Immunodeficiency Virus

Elena Chertova,1* Julian W. Bess, Jr.,1 Bruce J. Crise,1 Raymond C. Sowder II,1 Terra M. Schaden,1 Joanne M. Hilburn,1 James A. Hoxie,2 Raoul E. Benveniste,3 Jeffrey D. Lifson,1 Louis E. Henderson,1 and Larry O. Arthur1

AIDS Vaccine Program, SAIC Frederick,1 National Cancer Institute at Frederick, Frederick, Maryland 21702-1201,3 University of Pennsylvania, Philadelphia, Pennsylvania 191042

Received 10 October 2001/ Accepted 21 February 2002

Human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) particles typically contain small amounts of the surface envelope protein (SU), and this is widely believed to be due to shedding of SU from mature virions. We purified proteins from HIV-1 and SIV isolates using procedures which allow quantitative measurements of viral protein content and determination of the ratios of gag- and env-encoded proteins in virions. All of the HIV-1 and most of the SIV isolates examined contained low levels of envelope proteins, with Gag:Env ratios of approximately 60:1. Based on an estimate of 1,200 to 2,500 Gag molecules per virion, this corresponds to an average of between 21 and 42 SU molecules, or between 7 and 14 trimers, per particle. In contrast, some SIV isolates contained levels of SU at least 10-fold greater than SU from HIV-1 isolates. Quantification of relative amounts of SU and transmembrane envelope protein (TM) provides a means to assess the impact of SU shedding on virion SU content, since such shedding would be expected to result in a molar excess of TM over SU on virions that had shed SU. With one exception, viruses with sufficient SU and TM to allow quantification were found to have approximately equivalent molar amounts of SU and TM. The quantity of SU associated with virions and the SU:TM ratios were not significantly changed during multiple freeze-thaw cycles or purification through sucrose gradients. Exposure of purified HIV-1 and SIV to temperatures of 55°C or greater for 1 h resulted in loss of most of the SU from the virus but retention of TM. Incubation of purified virus with soluble CD4 at 37°C resulted in no appreciable loss of SU from either SIV or HIV-1. These results indicate that the association of SU and TM on the purified virions studied is quite stable. These findings suggest that incorporation of SU-TM complexes into the viral membrane may be the primary factor determining the quantity of SU associated with SIV and HIV-1 virions, rather than shedding of SU from mature virions.

* Corresponding author. Mailing address: National Cancer Institute at Frederick, Frederick, MD 21702-1201. Phone: (301) 846-1455. Fax: (301) 846-5588. E-mail: chertova{at}mail.ncifcrf.gov.

Journal of Virology, June 2002, p. 5315-5325, Vol. 76, No. 11
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.11.5315-5325.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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