Vet World Vol.17 December-2024 Article - 5
Research Article
Veterinary World, 17(12): 2736-2746
https://doi.org/10.14202/vetworld.2024.2736-2746
Benzimidazole resistance-associated mutations improve the in silico dimerization of hookworm tubulin: An additional resistance mechanism
2. Department of Veterinary Paraclinical Sciences, College of Veterinary Medicine, University of Southern Mindanao, Kabacan 9407, Cotabato, Philippines.
3. Department of Tropical Medicine, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand.
4. WHO Collaborating Center for Research and Control of Opisthorchiasis (Southeast Asian Liver Fluke Disease), Tropical Disease Research Center, Khon Kaen University, Khon Kaen 40002, Thailand.
Background and Aim: Mutations in the β-tubulin genes of helminths confer benzimidazole (BZ) resistance by reducing the drug’s binding efficiency to the expressed protein. However, the effects of these resistance-associated mutations on tubulin dimer formation in soil-transmitted helminths remain unknown. Therefore, this study aimed to investigate the impact of these mutations on the in silico dimerization of hookworm α- and β-tubulins using open-source bioinformatics tools.
Materials and Methods: Using AlphaFold 3, the α- and β-tubulin amino acid sequences of Ancylostoma ceylanicum were used to predict the structural fold of the hookworm tubulin heterodimer. The modeled complexes were subjected to several protein structure quality assurance checks. The binding free energies, overall binding affinity, dissociation constant, and interacting amino acids of the complex were determined. The dimer’s structural flexibility and motion were simulated through molecular dynamics.
Results: BZ resistance-associated amino acid substitutions in the β-tubulin isotype 1 protein of hookworms altered tubulin dimerization. The E198K, E198V, and F200Y mutations conferred the strongest and most stable binding between the α and β subunits, surpassing that of the wild-type. In contrast, complexes with the Q134H and F200L mutations exhibited the opposite effect. Molecular dynamics simulations showed that wild-type and mutant tubulin dimers exhibited similar dynamic behavior, with slight deviations in those carrying the F200L and E198K mutations.
Conclusion: Resistance-associated mutations in hookworms impair BZ binding to β-tubulin and enhance tubulin dimer interactions, thereby increasing the parasite’s ability to withstand treatment. Conversely, other mutations weaken these interactions, potentially compromising hookworm viability. These findings offer novel insights into helminth tubulin dimerization and provide a valuable foundation for developing anthelmintics targeting this crucial biological process.
Keywords: Ancylostoma, anthelmintic resistance, microtubules, soil-transmitted helminths.
How to cite this article: Tenorio JCB, Heikal MF, Kafle A, Saichua P, and Suttiprapa S (2024) Benzimidazole resistance-associated mutations improve the in silico dimerization of hookworm tubulin: An additional resistance mechanism, Veterinary World, 17(12): 2736-2746.
Received: 2024-08-16 Accepted: 2024-11-07 Published online: 2024-12-06
Corresponding author: E-mail:
DOI: 10.14202/vetworld.2024.2736-2746
Copyright: Tenorio, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/ by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
