Recombinant N-myristoyltransferase of Plasmodium falciparum (termed PfNMT) has been used in the development of a SPA (scintillation proximity assay) suitable for automation and high-throughput screening of inhibitors against this enzyme. The ability to use the SPA has been facilitated by development of an expression and purification system which yields considerably improved quantities of soluble active recombinant PfNMT compared with previous studies. Specifically, yields of pure protein have been increased from 12 μg·l−1 to >400 μg·l−1 by use of a synthetic gene with codon usage optimized for expression in an Escherichia coli host. Preliminary small-scale ‘piggyback’ inhibitor studies using the SPA have identified a family of related molecules containing a core benzothiazole scaffold with IC50 values <50 μM, which demonstrate selectivity over human NMT1. Two of these compounds, when tested against cultured parasites in vitro, reduced parasitaemia by >80% at a concentration of 10 μM.
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December 2007
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Research Article|
November 14 2007
Molecules incorporating a benzothiazole core scaffold inhibit the N-myristoyltransferase of Plasmodium falciparum
Paul W. Bowyer;
Paul W. Bowyer
1
*Wellcome Trust Laboratories for Molecular Parasitology, Imperial College London, London SW7 2AZ, U.K.
†Division of Cell and Molecular Biology, Centre for Molecular Microbiology and Infection, Imperial College London, London SW7 2AZ, U.K.
‡Department of Chemistry, Imperial College London, London SW7 2AZ, U.K.
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Ruwani S. Gunaratne;
Ruwani S. Gunaratne
2
§Division of Parasitology, MRC National Institute for Medical Research, Mill Hill, London NW7 1AA, U.K.
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Munira Grainger;
Munira Grainger
§Division of Parasitology, MRC National Institute for Medical Research, Mill Hill, London NW7 1AA, U.K.
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Chrislaine Withers-Martinez;
Chrislaine Withers-Martinez
§Division of Parasitology, MRC National Institute for Medical Research, Mill Hill, London NW7 1AA, U.K.
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Sasala R. Wickramsinghe;
Sasala R. Wickramsinghe
‡Department of Chemistry, Imperial College London, London SW7 2AZ, U.K.
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Edward W. Tate;
Edward W. Tate
‡Department of Chemistry, Imperial College London, London SW7 2AZ, U.K.
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Robin J. Leatherbarrow;
Robin J. Leatherbarrow
‡Department of Chemistry, Imperial College London, London SW7 2AZ, U.K.
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Katherine A. Brown;
Katherine A. Brown
†Division of Cell and Molecular Biology, Centre for Molecular Microbiology and Infection, Imperial College London, London SW7 2AZ, U.K.
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Anthony A. Holder;
Anthony A. Holder
3
§Division of Parasitology, MRC National Institute for Medical Research, Mill Hill, London NW7 1AA, U.K.
3Correspondence can be addressed to either of these authors (email dfs501@york.ac.uk or aholder@nimr.mrc.ac.uk).
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Deborah F. Smith
Deborah F. Smith
3
*Wellcome Trust Laboratories for Molecular Parasitology, Imperial College London, London SW7 2AZ, U.K.
†Division of Cell and Molecular Biology, Centre for Molecular Microbiology and Infection, Imperial College London, London SW7 2AZ, U.K.
∥Immunology and Infection Unit, Department of Biology/Hull York Medical School, University of York, Heslington, York YO10 5YW, U.K.
3Correspondence can be addressed to either of these authors (email dfs501@york.ac.uk or aholder@nimr.mrc.ac.uk).
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Publisher: Portland Press Ltd
Received:
May 24 2007
Revision Received:
August 14 2007
Accepted:
August 23 2007
Accepted Manuscript online:
August 23 2007
Online ISSN: 1470-8728
Print ISSN: 0264-6021
© The Authors Journal compilation © 2007 Biochemical Society
2007
Biochem J (2007) 408 (2): 173–180.
Article history
Received:
May 24 2007
Revision Received:
August 14 2007
Accepted:
August 23 2007
Accepted Manuscript online:
August 23 2007
Citation
Paul W. Bowyer, Ruwani S. Gunaratne, Munira Grainger, Chrislaine Withers-Martinez, Sasala R. Wickramsinghe, Edward W. Tate, Robin J. Leatherbarrow, Katherine A. Brown, Anthony A. Holder, Deborah F. Smith; Molecules incorporating a benzothiazole core scaffold inhibit the N-myristoyltransferase of Plasmodium falciparum. Biochem J 1 December 2007; 408 (2): 173–180. doi: https://doi.org/10.1042/BJ20070692
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