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Comparative structural, kinetic and inhibitor studies of Trypanosoma brucei trypanothione reductase with T. cruzi.
Mol Biochem Parasitol. 2010 Jan; 169(1):12-9.MB

Abstract

As part of a drug discovery programme to discover new treatments for human African trypanosomiasis, recombinant trypanothione reductase from Trypanosoma brucei has been expressed, purified and characterized. The crystal structure was solved by molecular replacement to a resolution of 2.3A and found to be nearly identical to the T. cruzi enzyme (root mean square deviation 0.6A over 482 Calpha atoms). Kinetically, the K(m) for trypanothione disulphide for the T. brucei enzyme was 4.4-fold lower than for T. cruzi measured by either direct (NADPH oxidation) or DTNB-coupled assay. The K(m) for NADPH for the T. brucei enzyme was found to be 0.77microM using an NADPH-regenerating system coupled to reduction of DTNB. Both enzymes were assayed for inhibition at their respective S=K(m) values for trypanothione disulphide using a range of chemotypes, including CNS-active drugs such as clomipramine, trifluoperazine, thioridazine and citalopram. The relative IC(50) values for the two enzymes were found to vary by no more than 3-fold. Thus trypanothione reductases from these species are highly similar in all aspects, indicating that they may be used interchangeably for structure-based inhibitor design and high-throughput screening.

Authors+Show Affiliations

The Wellcome Trust Biocentre, University of Dundee, Scotland, UK.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

19747949

Citation

Jones, Deuan C., et al. "Comparative Structural, Kinetic and Inhibitor Studies of Trypanosoma Brucei Trypanothione Reductase With T. Cruzi." Molecular and Biochemical Parasitology, vol. 169, no. 1, 2010, pp. 12-9.
Jones DC, Ariza A, Chow WH, et al. Comparative structural, kinetic and inhibitor studies of Trypanosoma brucei trypanothione reductase with T. cruzi. Mol Biochem Parasitol. 2010;169(1):12-9.
Jones, D. C., Ariza, A., Chow, W. H., Oza, S. L., & Fairlamb, A. H. (2010). Comparative structural, kinetic and inhibitor studies of Trypanosoma brucei trypanothione reductase with T. cruzi. Molecular and Biochemical Parasitology, 169(1), 12-9. https://doi.org/10.1016/j.molbiopara.2009.09.002
Jones DC, et al. Comparative Structural, Kinetic and Inhibitor Studies of Trypanosoma Brucei Trypanothione Reductase With T. Cruzi. Mol Biochem Parasitol. 2010;169(1):12-9. PubMed PMID: 19747949.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Comparative structural, kinetic and inhibitor studies of Trypanosoma brucei trypanothione reductase with T. cruzi. AU - Jones,Deuan C, AU - Ariza,Antonio, AU - Chow,Wing-Huen A, AU - Oza,Sandra L, AU - Fairlamb,Alan H, Y1 - 2009/09/10/ PY - 2009/06/17/received PY - 2009/08/27/revised PY - 2009/09/03/accepted PY - 2009/9/15/entrez PY - 2009/9/15/pubmed PY - 2010/1/26/medline SP - 12 EP - 9 JF - Molecular and biochemical parasitology JO - Mol Biochem Parasitol VL - 169 IS - 1 N2 - As part of a drug discovery programme to discover new treatments for human African trypanosomiasis, recombinant trypanothione reductase from Trypanosoma brucei has been expressed, purified and characterized. The crystal structure was solved by molecular replacement to a resolution of 2.3A and found to be nearly identical to the T. cruzi enzyme (root mean square deviation 0.6A over 482 Calpha atoms). Kinetically, the K(m) for trypanothione disulphide for the T. brucei enzyme was 4.4-fold lower than for T. cruzi measured by either direct (NADPH oxidation) or DTNB-coupled assay. The K(m) for NADPH for the T. brucei enzyme was found to be 0.77microM using an NADPH-regenerating system coupled to reduction of DTNB. Both enzymes were assayed for inhibition at their respective S=K(m) values for trypanothione disulphide using a range of chemotypes, including CNS-active drugs such as clomipramine, trifluoperazine, thioridazine and citalopram. The relative IC(50) values for the two enzymes were found to vary by no more than 3-fold. Thus trypanothione reductases from these species are highly similar in all aspects, indicating that they may be used interchangeably for structure-based inhibitor design and high-throughput screening. SN - 1872-9428 UR - https://www.unboundmedicine.com/medline/citation/19747949/Comparative_structural_kinetic_and_inhibitor_studies_of_Trypanosoma_brucei_trypanothione_reductase_with_T__cruzi_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-6851(09)00217-5 DB - PRIME DP - Unbound Medicine ER -