RESUMO
Peptides binding to a Gb3 mimic were selected from 12-mer peptide library. The self-assembled monolayer (SAM) of a Gb3 mimic was formed on the gold surface, and biopanning was carried out with the phage display peptide library. After three rounds of biopanning, four individual sequences were obtained from 10 phage clones, and the selected peptides having the specific 7-mer sequence (FHENWPS) showed affinities to the Gb3 mimic as strong as to RCA120. Molecular dynamics calculations suggested that the peptides bound to the Gb3 mimic by hydrophobic interaction and hydrogen bonding formation, and the cooperative interactions played an important role in the recognition. The Stx-1 binding was inhibited by the peptides.
Assuntos
Bacteriófagos/metabolismo , Mimetismo Molecular , Peptídeos/metabolismo , Triexosilceramidas/metabolismo , Sequência de Aminoácidos , Dicroísmo Circular , Ligação de Hidrogênio , Dados de Sequência Molecular , Ressonância Magnética Nuclear Biomolecular , Ligação Proteica , Triexosilceramidas/químicaRESUMO
Self-assembled monolayers (SAMs) of Gb3 mimics having different lengths of alkyl chains were prepared on gold surfaces, and their interactions with galactose-specific lectin (RCA(120)) and Shiga toxins (Stxs) were investigated by a quartz crystal microbalance (QCM) in aqueous solutions. Their interaction with RCA(120) was enhanced owing to the "cluster effect," regardless of the alkyl chain length of the SAMs. The interaction with Stxs was dependent on the alkyl chain length of Gb3 mimics. Stx-1 and Stx-2 showed a stronger affinity to the Gb3C2 SAM with ethyl disulfide and to the Gb3C10 with decyl disulfide, respectively. Gb3 glycoconjugate polymer with no alkyl spacer inhibited the adsorption of Stx-1 to Gb3C10 SAM but did not inhibit the adsorption of Stx-2 to Gb3C10 SAM. The results suggest that the alkyl chain of the glycolipid takes part in the binding to Stx-2 but not to Stx-1, which is also supported by the computer simulation of Stx-1 with a Gb3 model substance.