Rational engineering of a miniprotein that reproduces the core of the CD4 site interacting with HIV-1 envelope glycoprotein - Archive ouverte HAL Accéder directement au contenu
Article Dans Une Revue Proceedings of the National Academy of Sciences of the United States of America Année : 1999

Rational engineering of a miniprotein that reproduces the core of the CD4 site interacting with HIV-1 envelope glycoprotein

C. Vita
  • Fonction : Auteur
E. Drakopoulou
  • Fonction : Auteur
J. Vizzavona
  • Fonction : Auteur
Lilian Martin
C. Roumestand
  • Fonction : Auteur
Y.-S. Yang
  • Fonction : Auteur
L. Ylisastigui
  • Fonction : Auteur
A. Benjouad
  • Fonction : Auteur
J. Gluckman
  • Fonction : Auteur

Résumé

Protein-protein interacting surfaces are usually large and intricate, making the rational design of small mimetics of these interfaces a daunting problem. On the basis of a structural similarity between the CDR2-like loop of CD4 and the beta-hairpin region of a short scorpion toxin, scyllatoxin, we transferred the side chains of nine residues of CD4, central in the binding to HIV-1 envelope glycoprotein (gp120), to a structurally homologous region of the scorpion toxin scaffold. In competition experiments, the resulting 27-amino acid miniprotein inhibited binding of CD4 to gp120 with a 40 microM IC(50). Structural analysis by NMR showed that both the backbone of the chimeric beta-hairpin and the introduced side chains adopted conformations similar to those of the parent CD4. Systematic single mutations suggested that most CD4 residues from the CDR2-like loop were reproduced in the miniprotein, including the critical Phe-43. The structural and functional analysis performed suggested five additional mutations that, once incorporated in the miniprotein, increased its affinity for gp120 by 100-fold to an IC(50) of 0.1-1.0 microM, depending on viral strains. The resulting mini-CD4 inhibited infection of CD4(+) cells by different virus isolates. Thus, core regions of large protein-protein interfaces can be reproduced in miniprotein scaffolds, offering possibilities for the development of inhibitors of protein-protein interactions that may represent useful tools in biology and in drug discovery.

Dates et versions

hal-02528538 , version 1 (01-04-2020)

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Citer

C. Vita, E. Drakopoulou, J. Vizzavona, S. Rochette, Lilian Martin, et al.. Rational engineering of a miniprotein that reproduces the core of the CD4 site interacting with HIV-1 envelope glycoprotein. Proceedings of the National Academy of Sciences of the United States of America, 1999, 96 (23), pp.13091-13096. ⟨10.1073/pnas.96.23.13091⟩. ⟨hal-02528538⟩

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