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Teladorsagia circumcincta 1,6-Bisphosphate Aldolase: Molecular and biochemical characterisation, structure analysis and recognition by immune hosts

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posted on 2023-05-03, 21:18 authored by Saleh UmairSaleh Umair, Charlotte BouchetCharlotte Bouchet, Nik PalevichNik Palevich, Heather Simpson
A 1095 bp full length cDNA encoding Teladorsagia circumcincta aldolase (TciALDO-1) was cloned and expressed in Escherichia coli. Recombinant TciALDO-1 was purified, and its kinetic properties determined. The predicted protein consisted of 365 amino acids, and was present as a single band of about 44 kDa on SDS-PAGE. Multiple alignments of the protein sequence of TciALDO-1 with homologues from other helminths showed the greatest similarity (93%) to the aldolases of Haemonchus contortus and Dictyocaulus viviparus, 82–86% similarity to the other nematode sequences, and 68–71% similarity to cestode and trematode enzymes. Substrate binding sites and conserved regions were identified, and were completely conserved in other homologues. At 30 °C, the optimum pH for TciALDO-1 activity was pH 7.5, the Vmax was 432 ± 23 nmol × min−1 × mg−1 protein, and the apparent Km for the substrate fructose 1,6-bisphosphate was 0.24 ± 0.01 µM (mean ± SEM, n = 3). Recombinant TciALDO-1 was recognized by antibodies in both serum and saliva from field-immune sheep in ELISA, however, that was not the case with nematode-naïve sheep. Teladorsagia circumcincta fructose 1,6-bisphosphate aldolase appears to have potential as a vaccine candidate to control this common sheep parasite.

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© 2021 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/).

Language

  • English

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  • No

Publisher

MDPI

Journal title

Parasitologia

Citation

Umair, S., Bouchet, C., Palevich, N., & Simpson, H. (2021). Teladorsagia circumcincta 1,6-Bisphosphate Aldolase: Molecular and biochemical characterisation, structure analysis and recognition by immune hosts. Parasitologia, 1(1), 1–11. doi:10.3390/parasitologia1010001

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