Journal article
Novel Scorpion Toxin ω-Buthitoxin-Hf1a Selectively Inhibits Calcium Influx via CaV3.3 and CaV3.2 and Alleviates Allodynia in a Mouse Model of Acute Postsurgical Pain
International Journal of Molecular Sciences, Vol.25 (9), pp.1-18
2024
PMCID: PMC11084959
PMID: 38731963
Abstract
Venom peptides have evolved to target a wide range of membrane proteins through diverse mechanisms of action and structures, providing promising therapeutic leads for diseases, including pain, epilepsy, and cancer, as well as unique probes of ion channel structure-function. In this work, a high-throughput FLIPR window current screening assay on T-type CaV3.2 guided the isolation of a novel peptide named ω-Buthitoxin-Hf1a from scorpion Hottentotta franzwerneri crude venom. At only 10 amino acid residues with one disulfide bond, it is not only the smallest venom peptide known to target T-type CaVs but also the smallest structured scorpion venom peptide yet discovered. Synthetic Hf1a peptides were prepared with C-terminal amidation (Hf1a-NH2) or a free C-terminus (Hf1a-OH). Electrophysiological characterization revealed Hf1a-NH2 to be a concentration-dependent partial inhibitor of CaV3.2 (IC50 = 1.18 μM) and CaV3.3 (IC50 = 0.49 μM) depolarized currents but was ineffective at CaV3.1. Hf1a-OH did not show activity against any of the three T-type subtypes. Additionally, neither form showed activity against N-type CaV2.2 or L-type calcium channels. The three-dimensional structure of Hf1a-NH2 was determined using NMR spectroscopy and used in docking studies to predict its binding site at CaV3.2 and CaV3.3. As both CaV3.2 and CaV3.3 have been implicated in peripheral pain signaling, the analgesic potential of Hf1a-NH2 was explored in vivo in a mouse model of incision-induced acute post-surgical pain. Consistent with this role, Hf1a-NH2 produced antiallodynia in both mechanical and thermal pain.
Details
- Title
- Novel Scorpion Toxin ω-Buthitoxin-Hf1a Selectively Inhibits Calcium Influx via CaV3.3 and CaV3.2 and Alleviates Allodynia in a Mouse Model of Acute Postsurgical Pain
- Authors
- Dan Wang (Corresponding Author) - Jiangsu UniversityVolker Herzig - University of the Sunshine Coast, Queensland, School of Science, Technology and EngineeringZoltan Dekan - The University of QueenslandK Rosengren - The University of QueenslandColton Payne - The University of QueenslandJiajie Zhuang - Jiangsu UniversityEmmanuel Bourinet - Université de MontpellierLotten Ragnarsson - The University of QueenslandPaul Alewood - The University of QueenslandRichard Lewis (Corresponding Author) - The University of Queensland
- Publication details
- International Journal of Molecular Sciences, Vol.25 (9), pp.1-18
- Publisher
- MDPI AG
- Date published
- 2024
- DOI
- 10.3390/ijms25094745
- ISSN
- 1422-0067
- PMID
- 38731963; PMC11084959
- Copyright note
- © 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
- Data Availability
- The NMR structure and chemical shift assignments for Hf1a were submitted to PDB and BMRB and given accession codes 9BFL and 31169, respectively.
- Grants
- Organisation Unit
- School of Science, Technology and Engineering; Centre for Bioinnovation
- Language
- English
- Record Identifier
- 991018698702621
- Output Type
- Journal article
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