Abstract
Different toxins acting on Kv1.3 channel have been isolated from animal venom. MeuKTX toxin from Mesobuthus eupeus phillipsi scorpion and shtx-k toxin from Stichodactyla haddoni sea anemone have been identified as two effective Kv1.3 channel blockers. In this work, we characterized the genomic organization of both toxins. MeuKTX gene contains one intron and two exons, similar to the most scorpion toxins. There are a few reports of genomic structure of sea anemone toxins acting on Kv channels. The sequence encoding mature peptide of shtx-k was located in an exon separated by an intron from the coding exon of the propeptide and signal region. In order to make a peptide with more affinity for Kv1.3 channel and greater stability, the shtx-k/ MeuKTX chimeric peptide was designed and constructed using splicing by overlap extension-PCR (SOE-PCR) method. MeuKTX, shtx-k, and shtx-k/MeuKTX were cloned and the expression of the soluble proteins in E. coli was determined. Molecular docking studies indicated more inhibitory effect of shtx-k/MeuKTX on Kv1.3 channel compared to shtx-k and MeuKTX toxins. Key amino acids binding channel from both toxins, also involved in interaction of chimeric peptide with channel. Our results showed that the fusion peptide, shtx-k/MeuKTX could be an effective agent to target Kv1.3 channel.
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References
Anangi R, Koshy S, Huq R, Beeton C, Chuang WJ, King GF (2012) Recombinant expression of margatoxin and agitoxin-2 in pichia pastoris: an efficient method for production of KV 1.3 channel blockers. PLoS ONE 7(12):e52965
Bajaj S, Han J (2019) Venom-derived peptide modulators of cation-selective channels: Friend, foe or frenemy. Frontiers in pharmacology, vol 10. Frontiers Media, Lausanne, p 58. https://doi.org/10.3389/fphar.2019.00058
Beeton C, Pennington MW, Norton RS (2011) Analogs of the sea anemone potassium channel blocker ShK for the treatment of autoimmune diseases. Inflamm Allergy 10(5):313–321. https://doi.org/10.2174/187152811797200641
Bergeron Z, Bingham J-P (2012) Scorpion toxins specific for potassium (K+) channels: a historical overview of peptide bioengineering. Toxins 4(11):1082–1119. https://doi.org/10.3390/toxins4111082
Chang SC, Galea CA, Leung EWW, Tajhya RB, Beeton C, Pennington MW, Norton RS (2012) Expression and isotopic labelling of the potassium channel blocker ShK toxin as a thioredoxin fusion protein in bacteria. Toxicon 60(5):840–850. https://doi.org/10.1016/j.toxicon.2012.05.017
Chen Z, Hu Y, Hong J, Hu J, Yang W, Xiang F, Yang F, Xie Z, Cao Z, Li W, Lin D, Wu Y (2015) Toxin acidic residue evolutionary function-guided design of de novo peptide drugs for the immunotherapeutic target, the Kv1.3 channel. Sci Rep 5(1):1–8. https://doi.org/10.1038/srep09881
Chen R, Robinson A, Gordon D, Chung SH (2011) Modeling the binding of three toxins to the voltage-gated potassium channel (Kv1.3). Biophys J 101(11):2652–2660. https://doi.org/10.1016/j.bpj.2011.10.029
Dai L, Wu JJ, Gu YH, Lan ZD, Ling MH, Chi CW (2000) Genomic organization of three novel toxins from the scorpion Buthus martensi Karsch that are active on potassium channels. Biochem J 346(3):805–809. https://doi.org/10.1042/bj3460805
Edwards W, Fung-Leung WP, Huang C, Chi E, Wu N, Liu Y, Maher MP, Bonesteel R, Connor J, Fellows R (2014) Targeting the ion channel Kv1.3 with scorpion venom peptides engineered for potency, selectivity, and half-life. J Biol Chem 289(33):22704–22714
Finol-Urdaneta RK, Belovanovic A, Micic-Vicovac M, Kinsella GK, McArthur JR, Al-Sabi A (2020) Marine toxins targeting KV1 channels: pharmacological tools and therapeutic scaffolds. Marine drugs, vol 18. MDPI AG, Basel, p 173
Gao B, Peigneur S, Tytgat J, Zhu S (2010) A potent potassium channel blocker from Mesobuthus eupeus scorpion venom. Enferm Infecc Microbiol Clin 28(3):1847–1853. https://doi.org/10.1016/j.biochi.2010.08.003
Gendeh GS, Chung MCM, Jeyaseelan K (1997b) Genomic structure of a potassium channel toxin from Heteractis magnifica. FEBS Lett 418(1–2):183–188. https://doi.org/10.1016/S0014-5793(97)01365-3
Gendeh GS, Young LC, De Medeiros CLC, Jeyaseelan K, Harvey AL, Chung MCM (1997a) A new potassium channel toxin from the sea anemone Heteractis magnifica: isolation, cDNA cloning, and functional expression. Biochemistry 36(38):11461–11471. https://doi.org/10.1021/bi970253d
Jiménez-Vargas JM, Possani LD, Luna-Ramírez K (2017) Arthropod toxins acting on neuronal potassium channels. Neuropharmacology, vol 127. Elsevier Ltd, Amsterdam, pp 139–160. https://doi.org/10.1016/j.neuropharm.2017.09.025
Kuyucak S, Norton RS (2014) Computational approaches for designing potent and selective analogs of peptide toxins as novel therapeutics. Future medicinal chemistry, vol 6. Future Science, London, pp 1645–1658. https://doi.org/10.4155/FMC.14.98
Kuzmenkov AI, Vassilevski AA, Kudryashova KS, Nekrasova OV, Peigneur S, Tytgat J, Feofanov AV, Kirpichnikov MP, Grishin EV (2015) Variability of potassium channel blockers in Mesobuthus eupeus scorpion venom with focus on Kv1. 1 an integrated transcriptomic and proteomic study. J Biol Chem 290(19):12195–12209
Jin L, Wu Y (2007) Molecular mechanism of the sea anemone toxin ShK recognizing the Kv1.3 channel explored by docking and molecular dynamic simulations. J Chem Inf Model 47(5):1967–1972. https://doi.org/10.1021/ci700178w
Long SB, Campbell EB, MacKinnon R (2005) Crystal structure of a mammalian voltage-dependent Shaker family K + channel. Science 309(5736):897–903. https://doi.org/10.1126/science.1116269
MacKinnon R (2004) Potassium channels and the atomic basis of selective ion conduction (nobel lecture). Angew Chem Int Ed 43(33):4265–4277. https://doi.org/10.1002/ANIE.200400662@10.1002/(ISSN)1521-3773.NOBELLECTURES
Moran Y, Weinberger H, Sullivan JC, Reitzel AM, Finnerty JR, Gurevitz M (2008) Concerted evolution of sea anemone neurotoxin genes is revealed through analysis of the Nematostella vectensis genome. Mol Biol Evol 25(4):737–747
Murray JK, Qian YX, Liu B, Elliott R, Aral J, Park C, Zhang X, Stenkilsson M, Salyers K, Rose M, Li H, Yu S, Andrews KL, Colombero A, Werner J, Gaida K, Sickmier EA, Miu P, Itano A, Miranda LP (2015) Pharmaceutical optimization of peptide toxins for ion channel targets: potent, selective, and long-lived antagonists of Kv1.3. J Med Chem 58(17):6784–6802. https://doi.org/10.1021/acs.jmedchem.5b00495
Orts DJB, Moran Y, Cologna CT, Peigneur S, Madio B, Praher D, Quinton L, De Pauw E, Bicudo JEPW, Tytgat J, de Freitas JC (2013) BcsTx3 is a founder of a novel sea anemone toxin family of potassium channel blocker. FEBS J 280(19):4839–4852. https://doi.org/10.1111/febs.12456
Renisio JG, RomiLebrun R, Blanc E, Bornet O, Nakajima T, Darbon H (2000) Solution structure of BmKTX, a K+ blocker toxin from the Chinese scorpion Buthus martensi. Proteins 38(1):70–78. https://doi.org/10.1002/(SICI)1097-0134(20000101)38:1%3c70::AID-PROT8%3e3.0.CO;2-5
Shen B, Cao Z, Li W, Sabatier JM, Wu Y (2017) Treating autoimmune disorders with venom-derived peptides. Expert opinion on biological therapy, vol 17. Taylor and Francis Ltd, Oxfordshire, pp 1065–1075. https://doi.org/10.1080/14712598.2017.1346606
Tajti G, Wai DCC, Panyi G, Norton RS (2020) The voltage-gated potassium channel KV1.3 as a therapeutic target for venom-derived peptides. Biochemical pharmacology, vol 181. Elsevier Inc, Amsterdam, p 114146. https://doi.org/10.1016/j.bcp.2020.114146
Tudor JE, Pallaghy PK, Pennington MW, Norton RS (1996) Solution structure of ShK toxin, a novel potassium channel inhibitor from a sea anemone. Nat Struct Biol 3(4):317–320
Novoseletsky VN, Volyntseva AD, Shaitan KV, Kirpichnikov MP, Feofanov AV (2016) Modeling of the binding of peptide blockers to voltage-gated potassium channels: approaches and evidence. Acta Nat 8(2):35–46. https://doi.org/10.32607/20758251-2016-8-2-35-46
Varga Z, Tajti G, Panyi G (2021) The Kv1.3 K+ channel in the immune system and its “precision pharmacology” using peptide toxins. Biologia futura, vol 72. Akademiai Kiado ZRt, Budapest, pp 75–83. https://doi.org/10.1007/s42977-021-00071-7
Wang X, Li G, Guo J, Zhang Z, Zhang S, Zhu Y, Cheng J, Yu L, Ji Y, Tao J (2020) Kv1.3 channel as a key therapeutic target for neuroinflammatory diseases: state of the art and beyond. Frontiers in neuroscience, vol 13. Frontiers Media SA, Lausanne, p 1393. https://doi.org/10.3389/fnins.2019.01393
Wu S, Nie Y, Zeng XC, Cao H, Zhang L, Zhou L, Yang Y, Luo X, Liu Y (2014) Genomic and functional characterization of three new venom peptides from the scorpion Heterometrus spinifer. Peptides 53:30–41. https://doi.org/10.1016/j.peptides.2013.12.012
Wu Y, Wang L, Zhou M, You Y, Zhu X, Qiang Y, Qin M, Luo S, Ren Z, Xu A (2013) Correction: molecular evolution and diversity of conus peptide toxins, as revealed by gene structure and intron sequence analyses. PLoS ONE. https://doi.org/10.1371/annotation/98e70cd9-f5d7-4937-bdbc-c68bde86e8cf
Wulff H, Castle NA, Pardo LA (2009) Voltage-gated potassium channels as therapeutic targets. Nature reviews drug discovery, vol 8. NIH Public Access, Bethesda, pp 982–1001. https://doi.org/10.1038/nrd2983
Zhao Y, Huang J, Yuan X, Peng B, Liu W, Han S, He X (2015) Toxins targeting the KV1.3 channel: potential immunomodulators for autoimmune diseases. Toxins 7(5):1749–1764. https://doi.org/10.3390/toxins7051749
Zhou Q, Wang QL, Meng X, Shu Y, Jiang T, Wagenknecht T, Yin CC, Sui SF, Liu Z (2008) Structural and functional characterization of ryanodine receptor-natrin toxin interaction. Biophys J 95(9):4289–4299. https://doi.org/10.1529/biophysj.108.137224
Zhu S, Gao B, Aumelas A, del Carmen Rodríguez M, Lanz-Mendoza H, Peigneur S, Diego-Garcia E, Martin-Eauclaire MF, Tytgat J, Possani LD (2010) MeuTXKβ1, a scorpion venom-derived two-domain potassium channel toxin-like peptide with cytolytic activity. Biochim Et Biophys Acta 1804(4):872–883. https://doi.org/10.1016/j.bbapap.2009.12.017
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The authors would like to thank Shahrekord University, for providing the necessary equipment and materials.
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This work was supported by Shahrekord University.
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All authors contributed to the study conception and design. Material preparation was performed by HA. The biological experiments and data collection were done by MA. The first draft of the manuscript was written by MA and all authors commented on previous versions of the manuscript. HA as a supervisor of the project and corresponding author, and AMA and MSR as advisers of the study, designed the whole experiments, supported the project, analyzed data, and wrote the final manuscript. All authors read and approved the final manuscript.
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Asadi, M., Ayat, H., Ahadi, A.M. et al. Genomic Structure of Two Kv1.3 Channel Blockers from Scorpion Mesobuthus eupeus and Sea Anemone Stichodactyla haddoni and Construction of their Chimeric Peptide as a Novel Blocker. Biochem Genet 60, 504–526 (2022). https://doi.org/10.1007/s10528-021-10109-z
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DOI: https://doi.org/10.1007/s10528-021-10109-z