Artificial pore blocker acts specifically on voltage-gated potassium channel isoform K_V1.6

Among voltage-gated potassium channel (K_V) isoforms, K_V1.6 is one of the most widespread in the nervous system. However, there are little data concerning its physiological significance, in part due to the scarcity of specific ligands. The known high-affinity ligands of K_V1.6 lack selectivity, and conversely, its selective ligands show low affinity. Here, we present a designer peptide with both high affinity and selectivity to K_V1.6. Previously, we have demonstrated that K_V isoform-selective peptides can be constructed based on the simplistic α-hairpinin scaffold, and we obtained a number of artificial Tk-hefu peptides showing selective blockage of K_V1.3 in the submicromolar range. We have now proposed amino acid substitutions to enhance their activity. As a result, we have been able to produce Tk-hefu-11 that shows an EC₅₀ of ≈70 nM against K_V1.3. Quite surprisingly, Tk-hefu-11 turns out to block K_V1.6 with even higher potency, presenting an EC₅₀ of ≈10 nM. Furthermore, we have solved the peptide structure and used molecular dynamics to investigate the determinants of selective interactions between artificial α-hairpinins and K_V channels to explain the dramatic increase in K_V1.6 affinity. Since K_V1.3 is not highly expressed in the nervous system, we hope that Tk-hefu-11 will be useful in studies of K_V1.6 and its functions.