Frontiers in Cellular and Infection Microbiology (Aug 2024)

A Toxoplasma gondii thioredoxin with cell adhesion and antioxidant function

  • Dawei Wang,
  • Dawei Wang,
  • Yuyi Shi,
  • Yuyi Shi,
  • Ziwen Cheng,
  • Ziwen Cheng,
  • Like Luo,
  • Like Luo,
  • Kuo Cheng,
  • Kuo Cheng,
  • Shengqi Gan,
  • Shengqi Gan,
  • Che Liu,
  • Che Liu,
  • Zeliang Chen,
  • Baoling Yang,
  • Baoling Yang

DOI
https://doi.org/10.3389/fcimb.2024.1404120
Journal volume & issue
Vol. 14

Abstract

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BackgroundToxoplasma gondii (T. gondii) is a widespread, zoonotic protozoan intracellular parasite with a complex life cycle, which can cause toxoplasmosis, a potentially serious disease. During the invasion process, T. gondii proteins first bind to the relevant host cell receptors, such as glycosaminoglycan molecule (GAG-binding motif), which is one of the main receptors for parasites or virus to infect host cells. However, research on TGME49_216510 (T. gondii Trx21), a protein from Toxoplasma gondii, is limited.MethodsBioinformatics analysis of the Trx21 protein was performed firstly. And specific primers were then designed using the conserved domain and GAG-binding motif to amplify, express, and purify a fragment of the Trx21 protein. The purified Trx21-GST protein was used for antioxidant and cell adhesion experiments. Simultaneously, mice were immunized with Trx21-His to generate specific polyclonal antibodies for subcellular localization analysis.ResultsThe Trx21 protein, consisting of 774 amino acids, included a transmembrane region, three GAG-binding motifs, and a Thioredoxin-like domain. The recombinant Trx21-His protein had a molecular mass of about 31 kDa, while the Trx21-GST protein had a molecular mass of about 55 kDa, which was analyzed by SDS-PAGE and Western blot. Subcellular localization analysis by IFA revealed that Trx21 is predominantly distributed in the cytoplasm of T. gondii. Furthermore, Trx21 exhibited a protective effect on supercoiled DNA against metal-catalyzed oxidation (MCO) and demonstrated adhesion abilities to Vero cells.ConclusionsThese results indicate that Trx21 plays an important role in host cell interaction and oxidative damage.

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