Sequence-selective recognition of cationic amphipathic tripeptides with similar structures in aqueous solutions by cucurbit[7]uril

Sequence-selective recognition of cationic amphipathic peptides by synthetic receptors is significant to biological applications, but it is still a great challenging task. Here we first study the binding characteristics of receptor cucurbit[7]uril (CB[7]) to the smallest aromatic tripeptides X₁GG (X₁= tryptophan (W), phenylalanine (F), and tyrosine (Y)) and basic tripeptides X₂GG (X₂= arginine (R), lysine (K), and histidine (H)) by molecular dynamics simulations. The study indicates that the sidechains of aromatic X₁residues can be encapsulated into the CB[7] cavity, while the sidechains of basic X₂residues prefer to locate at the CB[7] portal. Based on that, we consider hydrophobic aromatic residues as the N-terminus, the smallest glycine (G) as the 2nd-residue and basic residues as the C-terminus, and design nine tripeptides X₁GX₂(X₁= F, Y, W and X₂= H, K, R). We found that there is a great influence of the C-terminal basic residue of X₁GX₂on binding with CB[7] due to the introduction of a new binding site between CB[7] and the sidechain of the C-terminal residue. Interestingly, CB[7] can differentiate WGR and WGK with similar structures efficiently because of their eight orders of magnitude difference in the association constant (K_a). Besides, for WGR, YGR, and YGK with a nanomolar binding affinity (K_a> 10⁹M⁻¹), on reversing the sequence order of the 2nd-residue and 3rd-residue, theirK_areduces by about at least 1000-fold, implying the sequence dependence of CB[7] on recognizing these tripeptides. These results predict the potential applications of CB[7] in recognizing cationic amphipathic peptides.