The carbohydrate at asparagine 386 on HIV-1 gp120 is not essential for protein folding and function but is involved in immune evasion

Rogier W. Sanders, Eelco van Anken, Alexei A. Nabatov, I. Marije Marije, Ilja Bontjer, Dirk Eggink, Mark Melchers, Els Busser, Martijn M. Dankers, Fedde Groot, Ineke Braakman, Ben Berkhout, William A. Paxton

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Abstract

Background: The HIV-1 envelope glycoprotein gp120, which mediates viral attachment to target cells, consists for ∼50% of sugar, but the role of the individual sugar chains in various aspects of gp120 folding and function is poorly understood. Here we studied the role of the carbohydrate at position 386. We identified a virus variant that had lost the 386 glycan in an evolution study of a mutant virus lacking the disulfide bond at the base of the V4 domain. Results: The 386 carbohydrate was not essential for folding of wt gp120. However, its removal improved folding of a gp120 variant lacking the 385-418 disulfide bond, suggesting that it plays an auxiliary role in protein folding in the presence of this disulfide bond. The 386 carbohydrate was not critical for gp120 binding to dendritic cells (DC) and DC-mediated HIV-1 transmission to T cells. In accordance with previous reports, we found that N386 was involved in binding of the mannose-dependent neutralizing antibody 2G12. Interestingly, in the presence of specific substitutions elsewhere in gp120, removal of N386 did not result in abrogation of 2G12 binding, implying that the contribution of N386 is context dependent. Neutralization by soluble CD4 and the neutralizing CD4 binding site (CD4BS) antibody b12 was significantly enhanced in the absence of the 386 sugar, indicating that this glycan protects the CD4BS against antibodies. Conclusion: The carbohydrate at position 386 is not essential for protein folding and function, but is involved in the protection of the CD4BS from antibodies. Removal of this sugar in the context of trimeric Env immunogens may therefore improve the elicitation of neutralizing CD4BS antibodies.

Original languageEnglish
Article number10
JournalRetrovirology
Volume5
DOIs
Publication statusPublished - Jan 31 2008

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Antibody Binding Sites
Immune Evasion
Asparagine
Protein Folding
HIV-1
Carbohydrates
Disulfides
Dendritic Cells
Polysaccharides
HIV Envelope Protein gp120
Viruses
Mannose
Neutralizing Antibodies
T-Lymphocytes

ASJC Scopus subject areas

  • Virology

Cite this

Sanders, R. W., van Anken, E., Nabatov, A. A., Marije, I. M., Bontjer, I., Eggink, D., ... Paxton, W. A. (2008). The carbohydrate at asparagine 386 on HIV-1 gp120 is not essential for protein folding and function but is involved in immune evasion. Retrovirology, 5, [10]. https://doi.org/10.1186/1742-4690-5-10

The carbohydrate at asparagine 386 on HIV-1 gp120 is not essential for protein folding and function but is involved in immune evasion. / Sanders, Rogier W.; van Anken, Eelco; Nabatov, Alexei A.; Marije, I. Marije; Bontjer, Ilja; Eggink, Dirk; Melchers, Mark; Busser, Els; Dankers, Martijn M.; Groot, Fedde; Braakman, Ineke; Berkhout, Ben; Paxton, William A.

In: Retrovirology, Vol. 5, 10, 31.01.2008.

Research output: Contribution to journalArticle

Sanders, RW, van Anken, E, Nabatov, AA, Marije, IM, Bontjer, I, Eggink, D, Melchers, M, Busser, E, Dankers, MM, Groot, F, Braakman, I, Berkhout, B & Paxton, WA 2008, 'The carbohydrate at asparagine 386 on HIV-1 gp120 is not essential for protein folding and function but is involved in immune evasion', Retrovirology, vol. 5, 10. https://doi.org/10.1186/1742-4690-5-10
Sanders, Rogier W. ; van Anken, Eelco ; Nabatov, Alexei A. ; Marije, I. Marije ; Bontjer, Ilja ; Eggink, Dirk ; Melchers, Mark ; Busser, Els ; Dankers, Martijn M. ; Groot, Fedde ; Braakman, Ineke ; Berkhout, Ben ; Paxton, William A. / The carbohydrate at asparagine 386 on HIV-1 gp120 is not essential for protein folding and function but is involved in immune evasion. In: Retrovirology. 2008 ; Vol. 5.
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abstract = "Background: The HIV-1 envelope glycoprotein gp120, which mediates viral attachment to target cells, consists for ∼50{\%} of sugar, but the role of the individual sugar chains in various aspects of gp120 folding and function is poorly understood. Here we studied the role of the carbohydrate at position 386. We identified a virus variant that had lost the 386 glycan in an evolution study of a mutant virus lacking the disulfide bond at the base of the V4 domain. Results: The 386 carbohydrate was not essential for folding of wt gp120. However, its removal improved folding of a gp120 variant lacking the 385-418 disulfide bond, suggesting that it plays an auxiliary role in protein folding in the presence of this disulfide bond. The 386 carbohydrate was not critical for gp120 binding to dendritic cells (DC) and DC-mediated HIV-1 transmission to T cells. In accordance with previous reports, we found that N386 was involved in binding of the mannose-dependent neutralizing antibody 2G12. Interestingly, in the presence of specific substitutions elsewhere in gp120, removal of N386 did not result in abrogation of 2G12 binding, implying that the contribution of N386 is context dependent. Neutralization by soluble CD4 and the neutralizing CD4 binding site (CD4BS) antibody b12 was significantly enhanced in the absence of the 386 sugar, indicating that this glycan protects the CD4BS against antibodies. Conclusion: The carbohydrate at position 386 is not essential for protein folding and function, but is involved in the protection of the CD4BS from antibodies. Removal of this sugar in the context of trimeric Env immunogens may therefore improve the elicitation of neutralizing CD4BS antibodies.",
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T1 - The carbohydrate at asparagine 386 on HIV-1 gp120 is not essential for protein folding and function but is involved in immune evasion

AU - Sanders, Rogier W.

AU - van Anken, Eelco

AU - Nabatov, Alexei A.

AU - Marije, I. Marije

AU - Bontjer, Ilja

AU - Eggink, Dirk

AU - Melchers, Mark

AU - Busser, Els

AU - Dankers, Martijn M.

AU - Groot, Fedde

AU - Braakman, Ineke

AU - Berkhout, Ben

AU - Paxton, William A.

PY - 2008/1/31

Y1 - 2008/1/31

N2 - Background: The HIV-1 envelope glycoprotein gp120, which mediates viral attachment to target cells, consists for ∼50% of sugar, but the role of the individual sugar chains in various aspects of gp120 folding and function is poorly understood. Here we studied the role of the carbohydrate at position 386. We identified a virus variant that had lost the 386 glycan in an evolution study of a mutant virus lacking the disulfide bond at the base of the V4 domain. Results: The 386 carbohydrate was not essential for folding of wt gp120. However, its removal improved folding of a gp120 variant lacking the 385-418 disulfide bond, suggesting that it plays an auxiliary role in protein folding in the presence of this disulfide bond. The 386 carbohydrate was not critical for gp120 binding to dendritic cells (DC) and DC-mediated HIV-1 transmission to T cells. In accordance with previous reports, we found that N386 was involved in binding of the mannose-dependent neutralizing antibody 2G12. Interestingly, in the presence of specific substitutions elsewhere in gp120, removal of N386 did not result in abrogation of 2G12 binding, implying that the contribution of N386 is context dependent. Neutralization by soluble CD4 and the neutralizing CD4 binding site (CD4BS) antibody b12 was significantly enhanced in the absence of the 386 sugar, indicating that this glycan protects the CD4BS against antibodies. Conclusion: The carbohydrate at position 386 is not essential for protein folding and function, but is involved in the protection of the CD4BS from antibodies. Removal of this sugar in the context of trimeric Env immunogens may therefore improve the elicitation of neutralizing CD4BS antibodies.

AB - Background: The HIV-1 envelope glycoprotein gp120, which mediates viral attachment to target cells, consists for ∼50% of sugar, but the role of the individual sugar chains in various aspects of gp120 folding and function is poorly understood. Here we studied the role of the carbohydrate at position 386. We identified a virus variant that had lost the 386 glycan in an evolution study of a mutant virus lacking the disulfide bond at the base of the V4 domain. Results: The 386 carbohydrate was not essential for folding of wt gp120. However, its removal improved folding of a gp120 variant lacking the 385-418 disulfide bond, suggesting that it plays an auxiliary role in protein folding in the presence of this disulfide bond. The 386 carbohydrate was not critical for gp120 binding to dendritic cells (DC) and DC-mediated HIV-1 transmission to T cells. In accordance with previous reports, we found that N386 was involved in binding of the mannose-dependent neutralizing antibody 2G12. Interestingly, in the presence of specific substitutions elsewhere in gp120, removal of N386 did not result in abrogation of 2G12 binding, implying that the contribution of N386 is context dependent. Neutralization by soluble CD4 and the neutralizing CD4 binding site (CD4BS) antibody b12 was significantly enhanced in the absence of the 386 sugar, indicating that this glycan protects the CD4BS against antibodies. Conclusion: The carbohydrate at position 386 is not essential for protein folding and function, but is involved in the protection of the CD4BS from antibodies. Removal of this sugar in the context of trimeric Env immunogens may therefore improve the elicitation of neutralizing CD4BS antibodies.

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