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Type I and type II fatty acid biosynthesis in Eimeria tenella: enoyl reductase activity and structure

Published online by Cambridge University Press:  13 August 2007

J. Z. LU ,
S. P. MUENCH ,
M. ALLARY ,
S. CAMPBELL ,
C. W. ROBERTS ,
E. MUI ,
R. L. McLEOD ,
D. W. RICE  and
S. T. PRIGGE
J. Z. LU
Affiliation:
Department of Molecular Microbiology & Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA
S. P. MUENCH
Affiliation:
Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, UK
M. ALLARY
Affiliation:
Department of Molecular Microbiology & Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA
S. CAMPBELL
Affiliation:
Strathclyde Institute of Biomedical Sciences, University of Strathclyde, Glasgow G4 0NR, UK
C. W. ROBERTS
Affiliation:
Strathclyde Institute of Biomedical Sciences, University of Strathclyde, Glasgow G4 0NR, UK
E. MUI
Affiliation:
Department of Ophthalmology and Visual Sciences, University of Chicago, Chicago, IL 60637, USA
R. L. McLEOD
Affiliation:
Department of Ophthalmology and Visual Sciences, University of Chicago, Chicago, IL 60637, USA Department of Pediatrics (Infectious Diseases), and Pathology and Committees on Genetics, Molecular Medicine and Immunology and the College, University of Chicago, Chicago, IL 60637, USA
D. W. RICE
Affiliation:
Department of Molecular Biology and Biotechnology, University of Sheffield, Sheffield S10 2TN, UK
S. T. PRIGGE*
Affiliation:
Department of Molecular Microbiology & Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA
*
*Corresponding author: Department of Molecular Microbiology & Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, USA. Tel: +1 443 287 4822. Fax: +1 410 955 0105. E-mail address: sprigge@jhsph.edu
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Summary

Apicomplexan parasites of the genus Eimeria are the major causative agent of avian coccidiosis, leading to high economic losses in the poultry industry. Recent results show that Eimeria tenella harbours an apicoplast organelle, and that a key biosynthetic enzyme, enoyl reductase, is located in this organelle. In related parasites, enoyl reductase is one component of a type II fatty acid synthase (FAS) and has proven to be an attractive target for antimicrobial compounds. We cloned and expressed the mature form of E. tenella enoyl reductase (EtENR) for biochemical and structural studies. Recombinant EtENR exhibits NADH-dependent enoyl reductase activity and is inhibited by triclosan with an IC50 value of 60 nm. The crystal structure of EtENR reveals overall similarity with other ENR enzymes; however, the active site of EtENR is unoccupied, a state rarely observed in other ENR structures. Furthermore, the position of the central beta-sheet appears to block NADH binding and would require significant movement to allow NADH binding, a feature not previously seen in the ENR family. We analysed the E. tenella genomic database for orthologues of well-characterized bacterial and apicomplexan FAS enzymes and identified 6 additional genes, suggesting that E. tenella contains a type II FAS capable of synthesizing saturated, but not unsaturated, fatty acids. Interestingly, we also identified sequences that appear to encode multifunctional type I FAS enzymes, a feature also observed in Toxoplasma gondii, highlighting the similarity between these apicomplexan parasites.

Keywords

ApicomplexaapicoplastEimeria tenellatype I fatty acid synthasetype II fatty acid synthaseenoyl (ACP) reductase
Type
Research Article
Information
Parasitology , Volume 134 , Issue 14 , December 2007 , pp. 1949 - 1962
Copyright
Copyright © Cambridge University Press 2007

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