Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping
The F1FO-ATP synthase is one of the enzymes that is essential to meet the energy requirement of both the proliferating aerobic and hypoxic dormant stages of the life cycle of mycobacteria. Most F-ATP synthases consume ATP in the α3:β3 headpiece to drive the γ subunit, which couples ATP cleavage with...
| Main Authors: | , , , , , , |
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| Formato: | Journal Article |
| Idioma: | English |
| Publicado: |
2016
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| Acceso en liña: | https://hdl.handle.net/10356/83414 http://hdl.handle.net/10220/41395 |
| _version_ | 1826112424401436672 |
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| author | Hotra, Adam Suter, Manuel Biuković, Goran Priya, Ragunathan Kundu, Subhashri Dick, Thomas Grüber, Gerhard |
| author2 | School of Biological Sciences |
| author_facet | School of Biological Sciences Hotra, Adam Suter, Manuel Biuković, Goran Priya, Ragunathan Kundu, Subhashri Dick, Thomas Grüber, Gerhard |
| author_sort | Hotra, Adam |
| collection | NTU |
| description | The F1FO-ATP synthase is one of the enzymes that is essential to meet the energy requirement of both the proliferating aerobic and hypoxic dormant stages of the life cycle of mycobacteria. Most F-ATP synthases consume ATP in the α3:β3 headpiece to drive the γ subunit, which couples ATP cleavage with proton pumping in the c ring of FO via the bottom of the γ subunit. ATPase-driven H+ pumping is latent in mycobacteria. The presence of a unique 14 amino acid residue loop of the mycobacterial γ subunit has been described and aligned in close vicinity to the c-ring loop Priya R et al. (2013) J Bioenerg Biomembr 45, 121-129 Here, we used inverted membrane vesicles (IMVs) of fast-growing Mycobacterium smegmatis and a variety of covalent and non-covalent inhibitors to characterize the ATP hydrolysis activity of the F-ATP synthase inside IMVs. These vesicles formed a platform to investigate the function of the unique mycobaterial γ loop by deleting the respective loop-encoding sequence (γ166–179) in the genome of M. smegmatis. ATP hydrolysis-driven H+ pumping was observed in IMVs containing the Δγ166–179 mutant protein but not for IMVs containing the wild-type F-ATP synthase. In addition, when compared to the wild-type enzyme, IMVs containing the Δγ166–179 mutant protein showed increased ATP cleavage and lower levels of ATP synthesis, demonstrating that the loop affects ATPase activity, ATPase-driven H+ pumping and ATP synthesis. These results further indicate that the loop may affect coupling of ATP hydrolysis and synthesis in a different mode. |
| first_indexed | 2024-10-01T03:06:47Z |
| format | Journal Article |
| id | ntu-10356/83414 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T03:06:47Z |
| publishDate | 2016 |
| record_format | dspace |
| spelling | ntu-10356/834142020-11-01T04:44:17Z Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping Hotra, Adam Suter, Manuel Biuković, Goran Priya, Ragunathan Kundu, Subhashri Dick, Thomas Grüber, Gerhard School of Biological Sciences School of Physical and Mathematical Sciences Interdisciplinary Graduate School (IGS) Bioenergetics FATP synthase The F1FO-ATP synthase is one of the enzymes that is essential to meet the energy requirement of both the proliferating aerobic and hypoxic dormant stages of the life cycle of mycobacteria. Most F-ATP synthases consume ATP in the α3:β3 headpiece to drive the γ subunit, which couples ATP cleavage with proton pumping in the c ring of FO via the bottom of the γ subunit. ATPase-driven H+ pumping is latent in mycobacteria. The presence of a unique 14 amino acid residue loop of the mycobacterial γ subunit has been described and aligned in close vicinity to the c-ring loop Priya R et al. (2013) J Bioenerg Biomembr 45, 121-129 Here, we used inverted membrane vesicles (IMVs) of fast-growing Mycobacterium smegmatis and a variety of covalent and non-covalent inhibitors to characterize the ATP hydrolysis activity of the F-ATP synthase inside IMVs. These vesicles formed a platform to investigate the function of the unique mycobaterial γ loop by deleting the respective loop-encoding sequence (γ166–179) in the genome of M. smegmatis. ATP hydrolysis-driven H+ pumping was observed in IMVs containing the Δγ166–179 mutant protein but not for IMVs containing the wild-type F-ATP synthase. In addition, when compared to the wild-type enzyme, IMVs containing the Δγ166–179 mutant protein showed increased ATP cleavage and lower levels of ATP synthesis, demonstrating that the loop affects ATPase activity, ATPase-driven H+ pumping and ATP synthesis. These results further indicate that the loop may affect coupling of ATP hydrolysis and synthesis in a different mode. NMRC (Natl Medical Research Council, S’pore) MOH (Min. of Health, S’pore) Accepted version 2016-08-29T08:47:45Z 2019-12-06T15:21:59Z 2016-08-29T08:47:45Z 2019-12-06T15:21:59Z 2016 Journal Article Hotra, A., Suter, M., Biuković, G., Priya, R., Kundu, S., Dick, T., et al. (2016). Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping. FEBS Journal, 283(10), 1947-1961. 1742-464X https://hdl.handle.net/10356/83414 http://hdl.handle.net/10220/41395 10.1111/febs.13715 en FEBS Journal © 2016 Federation of European Biochemical Societies. This is the author created version of a work that has been peer reviewed and accepted for publication in FEBS Journal, published by John Wiley & Sons, Inc. on behalf of Federation of European Biochemical Societies. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at: [http://dx.doi.org/10.1111/febs.13715]. 64 p. application/pdf |
| spellingShingle | Bioenergetics FATP synthase Hotra, Adam Suter, Manuel Biuković, Goran Priya, Ragunathan Kundu, Subhashri Dick, Thomas Grüber, Gerhard Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping |
| title | Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping |
| title_full | Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping |
| title_fullStr | Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping |
| title_full_unstemmed | Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping |
| title_short | Deletion of a unique loop in the mycobacterial F-ATP synthase γ subunit sheds light on its inhibitory role in ATP hydrolysis-driven H+ pumping |
| title_sort | deletion of a unique loop in the mycobacterial f atp synthase γ subunit sheds light on its inhibitory role in atp hydrolysis driven h pumping |
| topic | Bioenergetics FATP synthase |
| url | https://hdl.handle.net/10356/83414 http://hdl.handle.net/10220/41395 |
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