Babesia bovis Dihydroorotate Dehydrogenase (BboDHODH) is a          Novel Molecular Target of Drug for Bovine Babesiosis

Ketsarin KAMYINGKIRD; Shinuo CAO; Tatsunori MASATANI; Paul Franck Adjou MOUMOUNI; Patrick VUDRIKO; Ahmed Abd El Moniem MOUSA; Mohamad Alaa TERKAWI; Yoshifumi NISHIKAWA; Ikuo IGARASHI; Xuenan XUAN

doi:10.1292/jvms.13-0419

Parasitology

Babesia bovis Dihydroorotate Dehydrogenase (BboDHODH) is a Novel Molecular Target of Drug for Bovine Babesiosis

Ketsarin KAMYINGKIRD, Shinuo CAO, Tatsunori MASATANI, Paul Franck Adjou MOUMOUNI, Patrick VUDRIKO, Ahmed Abd El Moniem MOUSA, Mohamad Alaa TERKAWI, Yoshifumi NISHIKAWA, Ikuo IGARASHI, Xuenan XUAN

Author information

Ketsarin KAMYINGKIRD
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Shinuo CAO
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Tatsunori MASATANI
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Paul Franck Adjou MOUMOUNI
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Patrick VUDRIKO
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Ahmed Abd El Moniem MOUSA
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Mohamad Alaa TERKAWI
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Yoshifumi NISHIKAWA
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Ikuo IGARASHI
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan
Xuenan XUAN
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan

Keywords: atovaquone, Babesia bovis, chemotherapeutic target agent, dihydroorotate dehydrogenase, leflunomide

JOURNAL FREE ACCESS

2014 Volume 76 Issue 3 Pages 323-330

DOI https://doi.org/10.1292/jvms.13-0419

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Abstract

The emergence of drug resistance and adverse side effects of current bovine babesiosis treatment suggest that the search of new drug targets and development of safer and effective compounds are required. This study focuses on dihydroorotate dehydrogenase (DHODH), the fourth enzyme of pyrimidine biosynthesis pathway as a potential drug target for bovine babesiosis. Recombinant Babesia bovis DHODH protein (rBboDHODH) was produced in Escherichia coli and used for characterization and measurement of enzymatic activity. Furthermore, the effects of DHODH inhibitors were evaluated in vitro. The recombinant B. bovis DHODH histidine fusion protein (rBboDHODH) had 42.4-kDa molecular weight and exhibited a specific activity of 475.7 ± 245 Unit/mg, a K_m = 276.2 µM for L-dihydroorotate and a K_m= 94.41 µM for decylubiquinone. A 44-kDa band of native BboDHODH was detected by Western blot analysis and found in parasites mitochondria using a confocal microscope. Among DHODH inhibitors, atovaquone (ATV) and leflunomide (LFN) significantly inhibited the activity of rBboDHODH as well as the growth of B. bovis in vitro. The half maximal inhibitory concentration (IC₅₀) of ATV and LFN was 2.38 ± 0.53 nM and 52.41 ± 11.47 µM, respectively. These results suggest that BboDHODH might be a novel target for development of new drug for treatment of B. bovis infection.

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