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Biochemical characteristics and inhibitor selectivity of mouse indoleamine 2,3-dioxygenase-2

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Abstract

The first step in the kynurenine pathway of tryptophan catabolism is the cleavage of the 2,3-double bond of the indole ring of tryptophan. In mammals, this reaction is performed independently by indoleamine 2,3-dioxygenase-1 (IDO1), tryptophan 2,3-dioxygenase (TDO) and the recently discovered indoleamine 2,3-dioxygenase-2 (IDO2). Here we describe characteristics of a purified recombinant mouse IDO2 enzyme, including its pH stability, thermal stability and structural features. An improved assay system for future studies of recombinant/isolated IDO2 has been developed using cytochrome b 5 as an electron donor. This, the first description of the interaction between IDO2 and cytochrome b 5, provides further evidence of the presence of a physiological electron carrier necessary for activity of enzymes in the “IDO family”. Using this assay, the kinetic activity and substrate range of IDO2 were shown to be different to those of IDO1. 1-Methyl-d-tryptophan, a current lead IDO inhibitor used in clinical trials, was a poor inhibitor of both IDO1 and IDO2 activity. This suggests that its immunosuppressive effect may be independent of pharmacological inhibition of IDO enzymes, in the mouse at least. The different biochemical characteristics of the mouse IDO proteins suggest that they have evolved to have distinct biological roles.

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Acknowledgments

This work was supported by the Australian Research Council, the National Health and Medical Research Council and the German Research Council (SFB 535, TP A12 to KB). Boniface Mailu is a scholar of the German Academic Exchange Service which is gratefully acknowledged. RS was supported by a NHMRC Senior Principal Research Fellowship, a University of Sydney Professorial Fellowship, and the University of Sydney Medical Foundation. We thank Joanne Jamie and Robert Willows of Macquarie University for use of the MUCAB CD spectrometer.

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Authors and Affiliations

  1. Molecular Immunopathology Unit, Discipline of Pathology and Bosch Institute, University of Sydney, Camperdown, NSW, 2006, Australia

    Christopher Jonathan Daraius Austin, A. Sanchez-Perez, N. H. Hunt & H. J. Ball

  2. Interdisciplinary Research Center, Giessen University, Giessen, Germany

    B. M. Mailu, S. Rahlfs, K. Zocher & K. Becker

  3. Centre for Vascular Research, School of Medical Sciences (Pathology) and Bosch Institute, Sydney Medical School, University of Sydney, Sydney, NSW, Australia

    G. J. Maghzal & R. Stocker

  4. Laboratory of Biochemistry, Department of Applied Science, Faculty of Science, National University Corporation Kochi University, Kochi, 780-8520, Japan

    H. J. Yuasa

  5. Discipline of Medicine, Central Clinical School, Sydney Bioinformatics, University of Sydney, Sydney, NSW, Australia

    J. W. Arthur

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  1. Christopher Jonathan Daraius Austin
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  2. B. M. Mailu
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  3. G. J. Maghzal
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  9. K. Becker
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  11. N. H. Hunt
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  12. H. J. Ball
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Correspondence to H. J. Ball.

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C. J. D. Austin and B. M. Mailu contributed equally to the work in this paper.

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Austin, C.J.D., Mailu, B.M., Maghzal, G.J. et al. Biochemical characteristics and inhibitor selectivity of mouse indoleamine 2,3-dioxygenase-2. Amino Acids 39, 565–578 (2010). https://doi.org/10.1007/s00726-010-0475-9

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