Novel miniproteins engineered by the transfer of active sites to small natural scaffolds.
Small multidisulfide-containing proteins are attractive structural templates to produce a biologically active conformation that mimics the binding surface of natural large proteins. In particular, the structural motif that is evolutionary conserved in all scorpion toxins has a small size (30-40 amino acid residues), a great structural stability, and high permissiveness for sequence mutation. This motif is composed of a beta-sheet and an alpha-helix bridged in the interior core by three disulfides. We have used this motif successfully to transfer within its beta-sheet new functional sites, including the curaremimetic loop of a snake neurotoxin and the CDR2-like site of human CD4. Accumulated evidence indicated that the two miniproteins produced, the curaremimetic miniprotein and the CD4 mimetic, contain the alpha/beta fold that is characteristic of the scaffold used and bind respectively to the acetylcholine receptor and to the envelope gp120 of HIV-1. Furthermore, the latter was shown to prevent viral infection of lymphocytes. These examples illustrate that, by the transfer of active sites to small and stable natural scaffolds, it is possible to engineer miniproteins reproducing, in part, the function of much larger proteins. Such miniproteins may be of great utility as tools in structure-function studies and as leads in drug design.
CEA Saclay, Département d'Ingénierie et d'Etudes des Protéines, Gif-sur-Yvette, France.
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Vita
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Vizzavona
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Drakopoulou
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Zinn-Justin
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Gilquin
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Ménez
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001704:Biopolymers / Q000737:chemistry; D015704:CD4 Antigens / Q000737:chemistry / Q000235:genetics; D003472:Curare / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000737:chemistry; D015195:Drug Design; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D015202:Protein Engineering; D011506:Proteins / Q000138:chemical synthesis* / Q000737:chemistry* / Q000235:genetics
D001704:Biopolymers; D015704:CD4 Antigens; D011506:Proteins; D003472:Curare