De novo designed library of linear helical peptides: an exploratory tool in the discovery of protein-protein interaction modulators.
Protein-protein interactions (PPIs) have emerged as important targets for pharmaceutical intervention because of their essential role in numerous physiological and pathological processes, but screening efforts using small-molecules have led to very low hit rates. Linear peptides could represent a quick and effective approach to discover initial PPI hits, particularly if they have inherent ability to adopt specific peptide secondary structures. Here, we address this hypothesis through a linear helical peptide library, composed of four sublibraries, which was designed by theoretical predictions of helicity (Agadir software). The 13-mer peptides of this collection fixes either a combination of three aromatic or two aromatic and one aliphatic residues on one face of the helix (Ac-SSEEX(5)ARNX(9)AAX(12)N-NH2), since these are structural features quite common at PPIs interfaces. The 81 designed peptides were conveniently synthesized by parallel solid-phase methodologies, and the tendency of some representative library components to adopt the intended secondary structure was corroborated through CD and NMR experiments. As proof of concept in the search for PPI modulators, the usefulness of this library was verified on the widely studied p53-MDM2 interaction and on the communication between VEGF and its receptor Flt-1, two PPIs for which a hydrophobic α-helix is essential for the interaction. We have demonstrated here that, in both cases, selected peptides from the library, containing the right hydrophobic sequence of the hot-spot in one of the protein partners, are able to interact with the complementary protein. Moreover, we have discover some new, quite potent inhibitors of the VEGF-Flt-1 interaction, just by replacing one of the aromatic residues of the initial F(5)Y(9)Y(12) peptide by W, in agreement with previous results on related antiangiogenic peptides. Finally, the HTS evaluation of the full collection on thermoTRPs has led to a few antagonists of TRPV1 and TRPA1 channels, which open new avenues on the way to innovative modulators of these channels.
Instituto de Química-Médica (IQM-CSIC) , Juan de la Cierva 3, 28006 Madrid, Spain.
M Ángeles
MÁ
Bonache
Beatriz
B
Balsera
Blanca
B
López-Méndez
Oscar
O
Millet
Diego
D
Brancaccio
Isabel
I
Gómez-Monterrey
Alfonso
A
Carotenuto
Luigi M
LM
Pavone
Marie
M
Reille-Seroussi
Nathalie
N
Gagey-Eilstein
Michel
M
Vidal
Roberto
R
de la Torre-Martinez
Asia
A
Fernández-Carvajal
Antonio
A
Ferrer-Montiel
M Teresa
MT
García-López
Mercedes
M
Martín-Martínez
M Jesús Pérez
MJ
de Vega
Rosario
R
González-Muñiz
D000595:Amino Acid Sequence; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D025941:Protein Interaction Mapping / Q000379:methods; D017433:Protein Structure, Secondary; D051736:Proto-Oncogene Proteins c-mdm2 / Q000378:metabolism; D060327:Solid-Phase Synthesis Techniques; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism; D040281:Vascular Endothelial Growth Factor Receptor-1 / Q000378:metabolism
D019151:Peptide Library; D010455:Peptides; D016159:Tumor Suppressor Protein p53; D042461:Vascular Endothelial Growth Factor A; D051736:Proto-Oncogene Proteins c-mdm2; D040281:Vascular Endothelial Growth Factor Receptor-1
NMR; P53-MDM2; TRPA1; ThermoTRPs; VEGF-Flt-1; peptides; protein−protein interactions; α-helix