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Journal of Molecular Evolution

, Volume 57, Issue 1, pp 1–15 | Cite as

The Evolution of Enzyme Specificity in Fasciola spp.

  • James A. Irving
  • Terry W. Spithill
  • Robert N. Pike
  • James C. Whisstock
  • Peter M. Smooker
Article

Abstract

Fasciola spp., commonly known as liver fluke, are significant trematode parasites of livestock and humans. They secrete several cathepsin L-like cysteine proteases, some of which differ in enzymatic properties and timing of expression in the parasite's life cycle. A detailed sequence and evolutionary analysis is presented, based on 18 cathepsin L-like enzymes isolated from Fasciola spp. (including a novel clone identified in this study). The enzymes form a monophyletic group which has experienced several gene duplication events over the last ~135 million years, giving rise to the present-day enzymatic repertoire of the parasite. This timing of these duplications appears to correlate with important points in the evolution of the mammalian hosts. Furthermore, the dates suggest that Fasciola hepatica and Fasciola gigantica diverged around 19 million years ago. A novel analysis, based on the pattern of amino acid diversity, was used to identify sites in the enzyme that are predicted to be subject to positive adaptive evolution. Many of these sites occur within the active site cleft of the enzymes, and hence would be expected to lead to differences in substrate specificity. Using homology modeling, with reference to previously obtained biochemical data, we are able to predict S2 subsite specificity for these enzymes: specifically those that can accommodate bulky hydrophobic residues in the P2 position and those that cannot. A number of other positions subject to evolutionary pressure and potentially significant for enzyme function are also identified, including sites anticipated to diminish cystatin binding affinity.

Keywords

Fasciola spp. Cathepsin L Enzyme specificity Phylogeny Molecular clock Adaptive evolution Sequence analysis 

Notes

Acknowledgements

The authors would like to thank Dr. David Blair for discussions, and Mr. Suharyanto for assistance with F. gigantica collection. JCW is a Monash University Logan Fellow and National Health and Medical Research Council Senior Research Fellow.

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Copyright information

© Springer-Verlag New York Inc. 2003

Authors and Affiliations

  • James A. Irving
    • 1
    • 2
  • Terry W. Spithill
    • 1
    • 3
  • Robert N. Pike
    • 1
  • James C. Whisstock
    • 1
    • 2
  • Peter M. Smooker
    • 4
  1. 1.Department of Biochemistry and Molecular BiologyMonash University, Victoria 3800Australia
  2. 2.Victorian Bioinformatics ConsortiumMonash University, Victoria 3800Australia
  3. 3.Institute of ParasitologyMcGill University, MontrealCanada
  4. 4.Department of Biotechnology and Environmental BiologyRMIT University, P.O. Box 71, Bundoora 3083Australia

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