Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalis
Acetyl phosphate (AcP) is generally produced from acetyl coenzyme A by phosphotransacetylase (Pta), and subsequent reaction with ADP, catalyzed by acetate kinase (Ack), produces ATP. The mechanism of ATP production in Porphyromonas gingivalis is poorly understood. The aim of this study was to explor...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2019-01-01
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| Series: | Journal of Oral Microbiology |
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| Online Access: | http://dx.doi.org/10.1080/20002297.2019.1588086 |
| _version_ | 1818855743864963072 |
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| author | Yasuo Yoshida Mitsunari Sato Takamasa Nonaka Yoshiaki Hasegawa Yuichiro Kezuka |
| author_facet | Yasuo Yoshida Mitsunari Sato Takamasa Nonaka Yoshiaki Hasegawa Yuichiro Kezuka |
| author_sort | Yasuo Yoshida |
| collection | DOAJ |
| description | Acetyl phosphate (AcP) is generally produced from acetyl coenzyme A by phosphotransacetylase (Pta), and subsequent reaction with ADP, catalyzed by acetate kinase (Ack), produces ATP. The mechanism of ATP production in Porphyromonas gingivalis is poorly understood. The aim of this study was to explore the molecular basis of the Pta-Ack pathway in this microorganism. Pta and Ack from P. gingivalis ATCC 33277 were enzymatically and structurally characterized. Structural and mutational analyses suggest that Pta is a dimer with two substrate-binding sites in each subunit. Ack is also dimeric, with a catalytic cleft in each subunit, and structural analysis indicates a dramatic domain motion that opens and closes the cleft during catalysis. ATP formation by Ack proceeds via a sequential mechanism. Reverse transcription-PCR analysis demonstrated that the pta (PGN_1179) and ack (PGN_1178) genes, tandemly located in the genome, are cotranscribed as an operon. Inactivation of pta or ack in P. gingivalis by homologous recombination was successful only when the inactivated gene was expressed in trans. Therefore, both pta and ack genes are essential for this microorganism. Insights into the Pta-Ack pathway reported herein would be helpful to understand the energy acquisition in P. gingivalis. |
| first_indexed | 2024-12-19T08:13:28Z |
| format | Article |
| id | doaj.art-e34c7a58bcfb4317baa86970c45873c0 |
| institution | Directory Open Access Journal |
| issn | 2000-2297 |
| language | English |
| last_indexed | 2024-12-19T08:13:28Z |
| publishDate | 2019-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Journal of Oral Microbiology |
| spelling | doaj.art-e34c7a58bcfb4317baa86970c45873c02022-12-21T20:29:33ZengTaylor & Francis GroupJournal of Oral Microbiology2000-22972019-01-0111110.1080/20002297.2019.15880861588086Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalisYasuo Yoshida0Mitsunari Sato1Takamasa Nonaka2Yoshiaki Hasegawa3Yuichiro Kezuka4School of Dentistry, Aichi Gakuin UniversitySchool of Dentistry, Aichi Gakuin UniversityIwate Medical UniversitySchool of Dentistry, Aichi Gakuin UniversityIwate Medical UniversityAcetyl phosphate (AcP) is generally produced from acetyl coenzyme A by phosphotransacetylase (Pta), and subsequent reaction with ADP, catalyzed by acetate kinase (Ack), produces ATP. The mechanism of ATP production in Porphyromonas gingivalis is poorly understood. The aim of this study was to explore the molecular basis of the Pta-Ack pathway in this microorganism. Pta and Ack from P. gingivalis ATCC 33277 were enzymatically and structurally characterized. Structural and mutational analyses suggest that Pta is a dimer with two substrate-binding sites in each subunit. Ack is also dimeric, with a catalytic cleft in each subunit, and structural analysis indicates a dramatic domain motion that opens and closes the cleft during catalysis. ATP formation by Ack proceeds via a sequential mechanism. Reverse transcription-PCR analysis demonstrated that the pta (PGN_1179) and ack (PGN_1178) genes, tandemly located in the genome, are cotranscribed as an operon. Inactivation of pta or ack in P. gingivalis by homologous recombination was successful only when the inactivated gene was expressed in trans. Therefore, both pta and ack genes are essential for this microorganism. Insights into the Pta-Ack pathway reported herein would be helpful to understand the energy acquisition in P. gingivalis.http://dx.doi.org/10.1080/20002297.2019.1588086porphyromonas gingivalisphosphotransacetylaseacetate kinaseatpessential genescrystal structure |
| spellingShingle | Yasuo Yoshida Mitsunari Sato Takamasa Nonaka Yoshiaki Hasegawa Yuichiro Kezuka Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalis Journal of Oral Microbiology porphyromonas gingivalis phosphotransacetylase acetate kinase atp essential genes crystal structure |
| title | Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalis |
| title_full | Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalis |
| title_fullStr | Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalis |
| title_full_unstemmed | Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalis |
| title_short | Characterization of the phosphotransacetylase-acetate kinase pathway for ATP production in Porphyromonas gingivalis |
| title_sort | characterization of the phosphotransacetylase acetate kinase pathway for atp production in porphyromonas gingivalis |
| topic | porphyromonas gingivalis phosphotransacetylase acetate kinase atp essential genes crystal structure |
| url | http://dx.doi.org/10.1080/20002297.2019.1588086 |
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