Abstract
Recombinant chimeras of hongotoxin 1 (HgTx1) fused through the N - or C-terminus with the red fluorescent protein TagRFP, RFP-HgTx1 and HgTx1-RFP were obtained. The properties of these fluorescent chimeras as ligands of potassium voltage-gated Kv1 channels were studied using bioengineered cell systems based on hybrid channels KcsA-Kv1.x (x = 1, 3, 6). It is shown that HgTx1-RFP and RFP-HgTx1 are high-affinity ligands of the external pore blocker binding sites of the Kv1.x (x = 1, 3) channels, and HgTx1-RFP is superior in activity to RFP-HgTx1. No specific interactions of HgTx1-RFP and RFP-HgTx1 with KcsA-Kv1. 6 were detected. It has been demonstrated that HgTx1-RFP and RFP-HgTx1 can be used in the KcsA-Kv1.x (x = 1, 3)-based cellular systems to search for Kv1.x (x = 1, 3) channel blockers and estimate their affinity.
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This work was supported by a grant from the Russian Science Foundation 19-74-30014.
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Abbreviations: AgTx2, agitoxin 2; GFP,green fluorescent protein; HgTx1, hongotoxin 1; HgTx1-RFP, hongotoxin 1 with the TagRFP protein fused at the C terminus; Kv-channels, potassium voltage-gated channels; Kv1.х (х = 1,3,6), potassium voltage-gated channels Kv1.1, Kv1.3, Kv1.6; MBP, maltose-binding protein; RFP, red fluorescence protein TagRFP; RFP-HgTx1, hongotoxin 1 with the TagRFP protein fused at the N terminus.
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Primak, A.L., Skutel, M.A., Nekrasova, O.V. et al. Kv1 Potassium Channel Ligands Based on Hongotoxin 1 and Red Fluorescent Protein. Russ J Bioorg Chem 46, 1011–1017 (2020). https://doi.org/10.1134/S1068162020060266
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DOI: https://doi.org/10.1134/S1068162020060266