AgResearch
Browse

File(s) not publicly available

Molecular and biochemical characterisation and recognition by the immune host of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) of the abomasal nematode parasite Teladorsagia circumcincta

journal contribution
posted on 2023-05-03, 16:43 authored by Saleh UmairSaleh Umair, Charlotte BouchetCharlotte Bouchet, Jacqui Knight, Tony Pernthaner, Heather Simpson
A 1023 bp full length cDNA encoding T. circumcincta GAPDH (TeciGAPDH) was cloned, expressed in Escherichia coli and the recombinant protein purified and its kinetic properties determined. A phylogenetic tree was constructed using helminth GAPDH sequences. The predicted protein consisted of 341 amino acids and was present as a single band of about 38 kDa on SDS-PAGE. Multiple alignments of the protein sequence of TeciGAPDH with homologues from other helminths showed that the greatest similarity (93%) to the GAPDH of Haemonchus contortus and Dictyocaulus viviparus, 82-96% 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 25 oC, the optimum pH for TeciGAPDH activity was pH 8, the Vmax was 1052 ± 23 nmoles.min-1.mg-1 protein and the Km for the substrate glyceraldehyde-3-phosphate was 0.02 ± 0.01 mM (both mean ± SD, n = 2). Antibodies in both serum and saliva from field-immune, but not nematode-naïve, sheep recognised recombinant TeciGAPDH in enzyme-linked immunosorbent assays. The recognition of the recombinant protein by antibodies generated by exposure of sheep to native enolase indicates similar antigenicity of the two proteins.

History

Rights statement

© 2017 Elsevier Inc. All rights reserved.

Language

  • English

Does this contain Māori information or data?

  • No

Publisher

Elsevier

Journal title

Experimental Parasitology

ISSN

0014-4894

Citation

Umair, S., Bouchet, C. L. G., Knight, J. S., Pernthaner, T., & Simpson, H. V. (2017). Molecular and biochemical characterisation and recognition by the immune host of the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) of the abomasal nematode parasite Teladorsagia circumcincta. Experimental Parasitology, 181, 40–46. doi:10.1016/j.exppara.2017.07.006

Funder

AGMARDT

Contract number

A20310

Job code

32758

Usage metrics

    Exports

    RefWorks
    BibTeX
    Ref. manager
    Endnote
    DataCite
    NLM
    DC