Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase
There is an alarming rate of human African trypanosomiasis recrudescence in many parts of sub-Saharan Africa. Yet, the disease has no successful chemotherapy. Trypanosoma lacks the enzymatic machinery for the de novo synthesis of purine nucleotides, and is critically dependent on salvage mechanisms....
| Main Authors: | , , , , , , , , , , , |
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| Format: | Journal Article |
| Language: | Inglés |
| Published: |
Cambridge University Press
2013
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10568/41905 |
| _version_ | 1855524240548167680 |
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| author | Tomoaki Bessho Shoko Morii Toshihide Kusumoto Takahiro Shinohara Masanori Noda Susumu Uchiyama Satoshi Shuto Shigenori Nishimura Duszenko, M. Martin, S.K. Takashi Inui Kubata, K.B. |
| author_browse | Duszenko, M. Kubata, K.B. Martin, S.K. Masanori Noda Satoshi Shuto Shigenori Nishimura Shoko Morii Susumu Uchiyama Takahiro Shinohara Takashi Inui Tomoaki Bessho Toshihide Kusumoto |
| author_facet | Tomoaki Bessho Shoko Morii Toshihide Kusumoto Takahiro Shinohara Masanori Noda Susumu Uchiyama Satoshi Shuto Shigenori Nishimura Duszenko, M. Martin, S.K. Takashi Inui Kubata, K.B. |
| author_sort | Tomoaki Bessho |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | There is an alarming rate of human African trypanosomiasis recrudescence in many parts of sub-Saharan Africa. Yet, the disease has no successful chemotherapy. Trypanosoma lacks the enzymatic machinery for the de novo synthesis of purine nucleotides, and is critically dependent on salvage mechanisms. Inosine 5′-monophosphate dehydrogenase (IMPDH) is responsible for the rate-limiting step in guanine nucleotide metabolism. Here, we characterize recombinant Trypanosoma brucei IMPDH (TbIMPDH) to investigate the enzymatic differences between TbIMPDH and host IMPDH. Size-exclusion chromatography and analytical ultracentrifugation sedimentation velocity experiments reveal that TbIMPDH forms a heptamer, different from type 1 and 2 mammalian tetrameric IMPDHs. Kinetic analysis reveals calculated K m values of 30 and 1300 μ m for IMP and NAD, respectively. The obtained K m value of TbIMPDH for NAD is approximately 20–200-fold higher than that of mammalian enzymes and indicative of a different NAD binding mode between trypanosomal and mammalian IMPDHs. Inhibition studies show K i values of 3·2 μ m, 21 nM and 3·3 nM for ribavirin 5′-monophosphate, mycophenolic acid and mizoribine 5′-monophosphate, respectively. Our results show that TbIMPDH is different from its mammalian counterpart and thus may be a good target for further studies on anti-trypanosomal drugs. |
| format | Journal Article |
| id | CGSpace41905 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2013 |
| publishDateRange | 2013 |
| publishDateSort | 2013 |
| publisher | Cambridge University Press |
| publisherStr | Cambridge University Press |
| record_format | dspace |
| spelling | CGSpace419052024-11-15T08:52:28Z Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase Tomoaki Bessho Shoko Morii Toshihide Kusumoto Takahiro Shinohara Masanori Noda Susumu Uchiyama Satoshi Shuto Shigenori Nishimura Duszenko, M. Martin, S.K. Takashi Inui Kubata, K.B. animal diseases animal health trypanosomiasis There is an alarming rate of human African trypanosomiasis recrudescence in many parts of sub-Saharan Africa. Yet, the disease has no successful chemotherapy. Trypanosoma lacks the enzymatic machinery for the de novo synthesis of purine nucleotides, and is critically dependent on salvage mechanisms. Inosine 5′-monophosphate dehydrogenase (IMPDH) is responsible for the rate-limiting step in guanine nucleotide metabolism. Here, we characterize recombinant Trypanosoma brucei IMPDH (TbIMPDH) to investigate the enzymatic differences between TbIMPDH and host IMPDH. Size-exclusion chromatography and analytical ultracentrifugation sedimentation velocity experiments reveal that TbIMPDH forms a heptamer, different from type 1 and 2 mammalian tetrameric IMPDHs. Kinetic analysis reveals calculated K m values of 30 and 1300 μ m for IMP and NAD, respectively. The obtained K m value of TbIMPDH for NAD is approximately 20–200-fold higher than that of mammalian enzymes and indicative of a different NAD binding mode between trypanosomal and mammalian IMPDHs. Inhibition studies show K i values of 3·2 μ m, 21 nM and 3·3 nM for ribavirin 5′-monophosphate, mycophenolic acid and mizoribine 5′-monophosphate, respectively. Our results show that TbIMPDH is different from its mammalian counterpart and thus may be a good target for further studies on anti-trypanosomal drugs. 2013-05 2014-08-06T11:56:46Z 2014-08-06T11:56:46Z Journal Article https://hdl.handle.net/10568/41905 en Limited Access Cambridge University Press Tomoaki Bessho, Shoko Morii, Toshihide Kusumoto, Takahiro Shinohara, Masanori Noda, Susumu Uchiyama, Satoshi Shuto, Shigenori Nishimura, Djikeng, A., Duszenko, M., Martin, S.K., Takashi Inui and Kubata, K.B. 2013. Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase. Parasitology 140(6):735-745. |
| spellingShingle | animal diseases animal health trypanosomiasis Tomoaki Bessho Shoko Morii Toshihide Kusumoto Takahiro Shinohara Masanori Noda Susumu Uchiyama Satoshi Shuto Shigenori Nishimura Duszenko, M. Martin, S.K. Takashi Inui Kubata, K.B. Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase |
| title | Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase |
| title_full | Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase |
| title_fullStr | Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase |
| title_full_unstemmed | Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase |
| title_short | Characterization of the novel Trypanosoma brucei inosine 5′-monophosphate dehydrogenase |
| title_sort | characterization of the novel trypanosoma brucei inosine 5 monophosphate dehydrogenase |
| topic | animal diseases animal health trypanosomiasis |
| url | https://hdl.handle.net/10568/41905 |
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