Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody - Archive ouverte HAL Access content directly
Journal Articles Retrovirology Year : 2014

Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody

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Julie Matz
  • Function : Author
  • PersonId : 930065
Marie-Christine Rouyez
  • Function : Author
  • PersonId : 955389
Annika Järviluoma
  • Function : Author
  • PersonId : 955390
Kalle Saksela
  • Function : Author
  • PersonId : 955391
Serge Benichou
  • Function : Author
  • PersonId : 955392

Abstract

Background
The HIV-1 Nef protein is essential for AIDS pathogenesis by its interaction with host cell surface receptors and signaling factors. Despite its critical role as a virulence factor Nef is not targeted by current antiviral strategies.
Results
We have determined the crystal structure of the complex formed by a camelid single-domain antibody fragment, termed sdAb19, bound to HIV-1 Nef together with a stabilizing SH3 domain. sdAb19 forms a stoichiometric 1:1 complex with Nef and binds to a conformationally conserved surface at the C-terminus of Nef that overlaps with functionally important interaction sites involved in Nef-induced perturbations of signaling and trafficking pathways. The antibody fragment binds Nef with low nanomolar affinity, which could be attenuated to micromolar affinity range by site-directed mutagenesis of key interaction residues in sdAb19. Fusion of the SH3 domain to sdAb19, termed Neffin, leads to a significantly increased affinity for Nef and formation of a stoichiometric 2:2 Nef-Neffin complex. The 19 kDa Neffin protein inhibits all functions of Nef as CD4 and MHC-I downregulation, association with Pak2, and the increase in virus infectivity and replication.
Conclusions
Together, sdAb19 and Neffin thus represent efficient tools for the rational development of antiviral strategies against HIV-1 Nef.
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Dates and versions

inserm-00980901 , version 1 (19-04-2014)

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Cite

Sebastian Lülf, Julie Matz, Marie-Christine Rouyez, Annika Järviluoma, Kalle Saksela, et al.. Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody. Retrovirology, 2014, 11 (1), pp.24. ⟨10.1186/1742-4690-11-24⟩. ⟨inserm-00980901⟩
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