1D5Q

SOLUTION STRUCTURE OF A MINI-PROTEIN REPRODUCING THE CORE OF THE CD4 SURFACE INTERACTING WITH THE HIV-1 ENVELOPE GLYCOPROTEIN

Summary for 1D5Q

• Entry DOI: 10.2210/pdb1d5q/pdb
• Related: 1CDH 1SCY
• Descriptor: CHIMERIC MINI-PROTEIN (1 entity in total)
• Functional Keywords: alpha-beta structure, charybdotoxin-like motif, binding protein
• Total number of polymer chains: 1
• Total formula weight: 2741.26

Authors

Vita, C.,Drakopoulou, E.,Vizzanova, J.,Rochette, S.,Martin, L.,Menez, A.,Roumestand, C.,Yang, Y.S.,Ylisastigui, L.,Benjouad, A.,Gluckman, J.C. (deposition date: 1999-10-11, release date: 2000-10-11, Last modification date: 2024-11-20)

Primary citation

Vita, C.,Drakopoulou, E.,Vizzavona, J.,Rochette, S.,Martin, L.,Menez, A.,Roumestand, C.,Yang, Y.S.,Ylisastigui, L.,Benjouad, A.,Gluckman, J.C.
Rational engineering of a miniprotein that reproduces the core of the CD4 site interacting with HIV-1 envelope glycoprotein.
Proc.Natl.Acad.Sci.USA, 96:13091-13096, 1999

PubMed Abstract: 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.

PubMed: 10557278
DOI: 10.1073/pnas.96.23.13091

Experimental method

SOLUTION NMR