Enhancing the Stability of Haemonchus contortus Glyceraldehyde-3-Phosphate Dehydrogenase and Binding of Host Albumin to the Parasite Enzyme

Abstract

Introduction

Haemonchus contortus is an economically important parasite of domestic animals. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) excreted in the ES product of H. contortus can be a promising vaccine candidate for controlling the parasite infection. Unfortunately, the parasite enzyme breaks down rapidly. The current study focusses on stabilizing the recombinant GAPDH (rGAPDH) of H. contortus.

Methods

The rGAPDH was purified and stored in two different buffers (sodium phosphate + EDTA and bicarbonate-sodium chloride) to check the stability. The affinity of the parasite enzyme towards host serum (Goat) components was evaluated by affinity chromatography. The interacting component was identified by mass spectrometry.

Results

Here, we report that the enzyme can be stabilized for at least 3 months if stored in bicarbonate-sodium chloride. This should facilitate testing of the enzyme in challenge trials. Additionally, we show that the parasite enzyme has affinity for host albumin; this interaction may have significance in host–parasite relationship.

Conclusion

The present study reports a combination of sodium bicarbonate (0.1 M) with 0.5 M sodium chloride as a suitable buffer to enhance the stability of H. contortus GAPDH.

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Acknowledgements

This work was supported by a Grant from the Department of Biotechnology, New Delhi, to P. J. YN was supported by IVRI and PKKM was recipient of ICMR fellowship. We thank Director, IVRI for providing the facilities.

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Contributions

PJ conceived the idea, partly executed, and wrote the manuscript. YN was involved in protein purification, stability testing, and analyzing GAPDH–albumin interactions, figure preparation, and reviewed the manuscript. PKKM was involved in blood collection, generation of affinity column, etc., and checked the draft.

Corresponding author

Correspondence to Prasanta Kumar K. Mishra.

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Nijo, Y., Mishra, P.K.K. & Joshi, P. Enhancing the Stability of Haemonchus contortus Glyceraldehyde-3-Phosphate Dehydrogenase and Binding of Host Albumin to the Parasite Enzyme. Acta Parasit. (2020). https://doi.org/10.2478/s11686-020-00212-3

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Keywords

  • Haemonchus contortus
  • GAPDH
  • Albumin
  • Stability
  • Buffer