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Design, expression, and immunogenicity of a soluble HIV trimeric envelope fragment adopting a prefusion gp41 configuration.

Publication ,  Journal Article
Qiao, Z-S; Kim, M; Reinhold, B; Montefiori, D; Wang, J-H; Reinherz, EL
Published in: J Biol Chem
June 17, 2005

The human immunodeficiency virus-1 (HIV-1) envelope glycoprotein (Env) is comprised of non-covalently associated gp120/gp41 subunits that form trimeric spikes on the virion surface. Upon binding to host cells, Env undergoes a series of structural transitions, leading to gp41 rearrangement necessary for fusion of viral and host membranes. Until now, the prefusion state of gp41 ectodomain (e-gp41) has eluded molecular and structural analysis, and thus assessment of the potential of such an e-gp41 conformer to elicit neutralizing antibodies has not been possible. Considering the importance of gp120 amino (C1) and carboxyl (C5) segments in the association with e-gp41, we hypothesize that these regions are sufficient to maintain e-gp41 in a prefusion state. Based on the available gp120 atomic structure, we designed several truncated gp140 variants by including the C1 and C5 regions of gp120 in a gp41 ectodomain fragment. After iterative cycles of protein design, expression and characterization, we obtained a variant truncated at Lys(665) that stably folds as an elongated trimer under physiologic conditions. Several independent biochemical/biophysical analyses strongly suggest that this mini-Env adopts a prefusion e-gp41 configuration that is strikingly distinct from the postfusion trimer-of-hairpin structure. Interestingly, this prefusion mini-Env, lacking the fragment containing the 2F5/4E10 neutralizing monoclonal antibody binding sites, displays no detectable HIV-neutralizing epitopes when employed as an immunogen in rabbits. The result of this immunogenicity study has important implications for HIV-1 vaccine design efforts. Moreover, this engineered mini-Env protein should facilitate three-dimensional structural studies of the prefusion e-gp41 and serve to guide future attempts at pharmacologic and immunologic intervention of HIV-1.

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Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

June 17, 2005

Volume

280

Issue

24

Start / End Page

23138 / 23146

Location

United States

Related Subject Headings

  • env Gene Products, Human Immunodeficiency Virus
  • Trypsin
  • Time Factors
  • Surface Plasmon Resonance
  • Scattering, Radiation
  • Rabbits
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Protein Engineering
  • Protein Conformation
 

Citation

APA
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ICMJE
MLA
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Qiao, Z.-S., Kim, M., Reinhold, B., Montefiori, D., Wang, J.-H., & Reinherz, E. L. (2005). Design, expression, and immunogenicity of a soluble HIV trimeric envelope fragment adopting a prefusion gp41 configuration. J Biol Chem, 280(24), 23138–23146. https://doi.org/10.1074/jbc.M414515200
Qiao, Zhi-Song, Mikyung Kim, Bruce Reinhold, David Montefiori, Jia-huai Wang, and Ellis L. Reinherz. “Design, expression, and immunogenicity of a soluble HIV trimeric envelope fragment adopting a prefusion gp41 configuration.J Biol Chem 280, no. 24 (June 17, 2005): 23138–46. https://doi.org/10.1074/jbc.M414515200.
Qiao Z-S, Kim M, Reinhold B, Montefiori D, Wang J-H, Reinherz EL. Design, expression, and immunogenicity of a soluble HIV trimeric envelope fragment adopting a prefusion gp41 configuration. J Biol Chem. 2005 Jun 17;280(24):23138–46.
Qiao, Zhi-Song, et al. “Design, expression, and immunogenicity of a soluble HIV trimeric envelope fragment adopting a prefusion gp41 configuration.J Biol Chem, vol. 280, no. 24, June 2005, pp. 23138–46. Pubmed, doi:10.1074/jbc.M414515200.
Qiao Z-S, Kim M, Reinhold B, Montefiori D, Wang J-H, Reinherz EL. Design, expression, and immunogenicity of a soluble HIV trimeric envelope fragment adopting a prefusion gp41 configuration. J Biol Chem. 2005 Jun 17;280(24):23138–23146.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

June 17, 2005

Volume

280

Issue

24

Start / End Page

23138 / 23146

Location

United States

Related Subject Headings

  • env Gene Products, Human Immunodeficiency Virus
  • Trypsin
  • Time Factors
  • Surface Plasmon Resonance
  • Scattering, Radiation
  • Rabbits
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Protein Engineering
  • Protein Conformation