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β-Sitosterol could serve as a dual inhibitor of Trypanosoma congolense sialidase and phospholipase A2: in vitro kinetic analyses and molecular dynamic simulations

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Abstract

The involvement of Trypanosoma congolense sialidase alongside phospholipase A2 has been widely accepted as the major contributing factor to anemia during African animal trypanosomiasis. The enzymes aid the parasite in scavenging sialic acid and fatty acids necessary for survival in the infected host, but there are no specific drug candidates against the two enzymes. This study investigated the inhibitory effects of β-sitosterol on the partially purified T. congolense sialidase and phospholipase A2. Purification of the enzymes using DEAE cellulose column led to fractions with highest specific activities of 8016.41 and 39.26 µmol/min/mg for sialidase and phospholipase A2, respectively. Inhibition kinetics studies showed that β-sitosterol is non-competitive and an uncompetitive inhibitor of sialidase and phospholipase A2 with inhibition binding constants of 0.368 and 0.549 µM, respectively. Molecular docking of the compound revealed binding energies of − 8.0 and − 8.6 kcal/mol against the sialidase and phospholipase A2, respectively. Furthermore, 100 ns molecular dynamics simulation using GROMACS revealed stable interaction of β-sitosterol with both enzymes. Hydrogen bond interactions between the ligand and Glu284 and Leu102 residues of the sialidase and phospholipase A2, respectively, were found to be the major stabilizing forces. In conclusion, β-sitosterol could serve as a dual inhibitor of T. congolense sialidase and phospholipase A2; hence, the compound could be exploited further in the search for newer trypanocides.

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Acknowledgements

We appreciate the management of Ahmadu Bello University, Zaria, Nigeria, for providing the enabling environment while carrying out the studies and a study fellowship to the first author. We also appreciate the support of the University of Groningen Peregrine for providing the computational resources for the molecular dynamic simulation studies. MAI is a recipient of the National Research Foundation Grant from TetFund, Nigeria (NRF2020/SETI/43). AUD is a beneficiary of The Ignacy Łukasiewicz Scholarship Programme from The Polish National Agency for Academic Exchange (NAWA).

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Funding was provided by Tertiary Education Trust Fund.

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Authors and Affiliations

  1. Department of Biochemistry, Ahmadu Bello University, Zaria, Nigeria

    Suleiman Aminu, Zainab Aliyu Alhafiz & Mohammed Auwal Ibrahim

  2. Biological and Chemical Research Center, Department of Chemistry, University of Warsaw, Warsaw, Poland

    Ammar Usman Danazumi & Maria Wiktoria Gorna

  3. Faculty of Chemistry, Warsaw University of Technology, Warsaw, Poland

    Ammar Usman Danazumi

  4. Department of Biochemistry, Federal University, Gusau, Nigeria

    Zainab Aliyu Alhafiz

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  1. Suleiman Aminu
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SA participated in the conceptualization of the study, methodology, resources, and initial draft of the manuscript. AUD wrote a section of the manuscript and performed the molecular dynamic simulation studies. ZAA was involved in formal analysis and editing. MWG was involved in formal analysis, investigation, writing—review, and editing. MAI contributed to the conceptualization of the study, methodology, formal analysis, data curation, supervision, writing—review, and editing.

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Aminu, S., Danazumi, A.U., Alhafiz, Z.A. et al. β-Sitosterol could serve as a dual inhibitor of Trypanosoma congolense sialidase and phospholipase A2: in vitro kinetic analyses and molecular dynamic simulations. Mol Divers 27, 1645–1660 (2023). https://doi.org/10.1007/s11030-022-10517-2

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