Structural Analysis of <i>Saccharomyces cerevisiae</i> Dihydroorotase Reveals Molecular Insights into the Tetramerization Mechanism
Dihydroorotase (DHOase), a dimetalloenzyme containing a carbamylated lysine within the active site, is a member of the cyclic amidohydrolase family, which also includes allantoinase (ALLase), dihydropyrimidinase (DHPase), hydantoinase, and imidase. Unlike most known cyclic amidohydrolases, which are...
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MDPI AG
2021-11-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/23/7249 |
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| author | Hong-Hsiang Guan Yen-Hua Huang En-Shyh Lin Chun-Jung Chen Cheng-Yang Huang |
| author_facet | Hong-Hsiang Guan Yen-Hua Huang En-Shyh Lin Chun-Jung Chen Cheng-Yang Huang |
| author_sort | Hong-Hsiang Guan |
| collection | DOAJ |
| description | Dihydroorotase (DHOase), a dimetalloenzyme containing a carbamylated lysine within the active site, is a member of the cyclic amidohydrolase family, which also includes allantoinase (ALLase), dihydropyrimidinase (DHPase), hydantoinase, and imidase. Unlike most known cyclic amidohydrolases, which are tetrameric, DHOase exists as a monomer or dimer. Here, we report and analyze two crystal structures of the eukaryotic <i>Saccharomyces cerevisiae</i> DHOase (ScDHOase) complexed with malate. The structures of different DHOases were also compared. An asymmetric unit of these crystals contained four crystallographically independent ScDHOase monomers. ScDHOase shares structural similarity with <i>Escherichia coli</i> DHOase (EcDHOase). Unlike EcDHOase, ScDHOase can form tetramers, both in the crystalline state and in solution. In addition, the subunit-interacting residues of ScDHOase for dimerization and tetramerization are significantly different from those of other DHOases. The tetramerization pattern of ScDHOase is also different from those of DHPase and ALLase. Based on sequence analysis and structural evidence, we identify two unique helices (α6 and α10) and a loop (loop 7) for tetramerization, and discuss why the residues for tetramerization in ScDHOase are not necessarily conserved among DHOases. |
| first_indexed | 2024-03-10T04:47:07Z |
| format | Article |
| id | doaj.art-2e408a7abcaf479495b6ba1a146e4878 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T04:47:07Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-2e408a7abcaf479495b6ba1a146e48782023-11-23T02:49:38ZengMDPI AGMolecules1420-30492021-11-012623724910.3390/molecules26237249Structural Analysis of <i>Saccharomyces cerevisiae</i> Dihydroorotase Reveals Molecular Insights into the Tetramerization MechanismHong-Hsiang Guan0Yen-Hua Huang1En-Shyh Lin2Chun-Jung Chen3Cheng-Yang Huang4Life Science Group, Scientific Research Division, National Synchrotron Radiation Research Center, Hsinchu 33076, TaiwanSchool of Biomedical Sciences, Chung Shan Medical University, No.110, Sec.1, Chien-Kuo N. Rd., Taichung City 402, TaiwanDepartment of Beauty Science, National Taichung University of Science and Technology, No.193, Sec.1, San-Min Rd., Taichung City 403, TaiwanLife Science Group, Scientific Research Division, National Synchrotron Radiation Research Center, Hsinchu 33076, TaiwanSchool of Biomedical Sciences, Chung Shan Medical University, No.110, Sec.1, Chien-Kuo N. Rd., Taichung City 402, TaiwanDihydroorotase (DHOase), a dimetalloenzyme containing a carbamylated lysine within the active site, is a member of the cyclic amidohydrolase family, which also includes allantoinase (ALLase), dihydropyrimidinase (DHPase), hydantoinase, and imidase. Unlike most known cyclic amidohydrolases, which are tetrameric, DHOase exists as a monomer or dimer. Here, we report and analyze two crystal structures of the eukaryotic <i>Saccharomyces cerevisiae</i> DHOase (ScDHOase) complexed with malate. The structures of different DHOases were also compared. An asymmetric unit of these crystals contained four crystallographically independent ScDHOase monomers. ScDHOase shares structural similarity with <i>Escherichia coli</i> DHOase (EcDHOase). Unlike EcDHOase, ScDHOase can form tetramers, both in the crystalline state and in solution. In addition, the subunit-interacting residues of ScDHOase for dimerization and tetramerization are significantly different from those of other DHOases. The tetramerization pattern of ScDHOase is also different from those of DHPase and ALLase. Based on sequence analysis and structural evidence, we identify two unique helices (α6 and α10) and a loop (loop 7) for tetramerization, and discuss why the residues for tetramerization in ScDHOase are not necessarily conserved among DHOases.https://www.mdpi.com/1420-3049/26/23/7249dihydroorotasetetramerizationpyrimidine biosynthesisCADdihydropyrimidinaseallantoinase |
| spellingShingle | Hong-Hsiang Guan Yen-Hua Huang En-Shyh Lin Chun-Jung Chen Cheng-Yang Huang Structural Analysis of <i>Saccharomyces cerevisiae</i> Dihydroorotase Reveals Molecular Insights into the Tetramerization Mechanism Molecules dihydroorotase tetramerization pyrimidine biosynthesis CAD dihydropyrimidinase allantoinase |
| title | Structural Analysis of <i>Saccharomyces cerevisiae</i> Dihydroorotase Reveals Molecular Insights into the Tetramerization Mechanism |
| title_full | Structural Analysis of <i>Saccharomyces cerevisiae</i> Dihydroorotase Reveals Molecular Insights into the Tetramerization Mechanism |
| title_fullStr | Structural Analysis of <i>Saccharomyces cerevisiae</i> Dihydroorotase Reveals Molecular Insights into the Tetramerization Mechanism |
| title_full_unstemmed | Structural Analysis of <i>Saccharomyces cerevisiae</i> Dihydroorotase Reveals Molecular Insights into the Tetramerization Mechanism |
| title_short | Structural Analysis of <i>Saccharomyces cerevisiae</i> Dihydroorotase Reveals Molecular Insights into the Tetramerization Mechanism |
| title_sort | structural analysis of i saccharomyces cerevisiae i dihydroorotase reveals molecular insights into the tetramerization mechanism |
| topic | dihydroorotase tetramerization pyrimidine biosynthesis CAD dihydropyrimidinase allantoinase |
| url | https://www.mdpi.com/1420-3049/26/23/7249 |
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