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Mechanistic and functional insights into fatty acid activation in Mycobacterium tuberculosis

Nature Chemical Biology volume 5, pages 166173 (2009) | Download Citation

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Abstract

The recent discovery of fatty acyl-AMP ligases (FAALs) in Mycobacterium tuberculosis (Mtb) provided a new perspective of fatty acid activation. These proteins convert fatty acids to the corresponding adenylates, which are intermediates of acyl-CoA–synthesizing fatty acyl-CoA ligases (FACLs). Presently, it is not evident how obligate pathogens such as Mtb have evolved such new themes of functional versatility and whether the activation of fatty acids to acyladenylates could indeed be a general mechanism. Here, based on elucidation of the first structure of an FAAL protein and by generating loss-of-function and gain-of-function mutants that interconvert FAAL and FACL activities, we demonstrate that an insertion motif dictates formation of acyladenylate. Because FAALs in Mtb are crucial nodes in the biosynthetic network of virulent lipids, inhibitors directed against these proteins provide a unique multipronged approach to simultaneously disrupting several pathways.

  • Compound

    Coenzyme A

  • Compound

    Adenosine 5′-triphosphate

  • Compound

    Lauroyl adenosine 5′-monophosphate

  • Compound

    Lauroyl coenzyme A

  • Compound

    5′-O-(N-Hexanoylsulfamoyl)adenosine

  • Compound

    5′-O-(N-Dodecanoylsulfamoyl)adenosine

  • Compound

    5′-O-(N-Icosanoylsulfamoyl)adenosine

  • Compound

    Methylmalonyl coenzyme A

  • Compound

    Adenosine

  • Compound

    2′,3′,4′-O,O,O-Tris(t-butyldimethylsilyl)adenosine

  • Compound

    2,′3′-Di-t-butyldimethylsilyl)adenosine

  • Compound

    2′,3′-O,O-Di-t-butyldimethylsilyl)-O-sulfamoyladenosine

  • Compound

    2′,3′-O,O-Di-t-butyldimethylsilyl)-O-(N-dodecanoyl)sulfamoyladenosine

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Acknowledgements

We thank L. Eltis (University of British Columbia) for Rhodococcus sp. RHA1 strain and S. Cole (Pasteur Research Institute, France) for the M. tuberculosis BAC genomic DNA library. P.A., A.G. and E.R. are Senior Research Fellows of the Council of Scientific and Industrial Research, India. R.S. is supported by a Wellcome Trust International Senior Research Fellowship in India. R.S.G. is supported by a Howard Hughes Medical Institute International Fellowship. This work is also partially supported by a Swarnajayanti Fellowship from the Department of Science and Technology of India and by a Centre of Excellence Grant from the Department of Biotechnology of India.

Author information

Author notes

    • Pooja Arora
    • , Aneesh Goyal
    •  & Vivek T Natarajan

    These authors contributed equally to this work.

Affiliations

  1. National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110 067, India.

    • Pooja Arora
    • , Vivek T Natarajan
    • , Priyanka Verma
    • , Omita A Trivedi
    • , Debasisa Mohanty
    •  & Rajesh S Gokhale
  2. Centre for Cellular and Molecular Biology, Council of Scientific and Industrial Research, Uppal Road, Hyderabad-500 007, India.

    • Aneesh Goyal
    • , Eerappa Rajakumara
    • , Malikmohamed Yousuf
    •  & Rajan Sankaranarayanan
  3. Department of Biochemistry, University of Delhi South Campus, Benito Juarez Road, New Delhi 110021, India.

    • Radhika Gupta
    •  & Anil Tyagi
  4. Jawaharlal Nehru Centre for Advanced Scientific Research, Jakkur, Bangalore 560064, India.

    • Rajesh S Gokhale

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Contributions

P.A., A.G., V.T.N., R.S. and R.S.G. designed the experiments, analyzed the data and wrote the manuscript. E.R. and M.Y. participated in structural studies. P.V., R.G. and O.A.T. participated in biochemical and mechanistic studies. D.M. and A.T. provided valuable advice.

Corresponding authors

Correspondence to Rajan Sankaranarayanan or Rajesh S Gokhale.

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    Supplementary Text and Figures

    Supplementary Figures 1–5, Supplementary Tables 1–4 and Supplementary Methods

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DOI

https://doi.org/10.1038/nchembio.143

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