Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/149231
Title: A systematic assessment of mycobacterial F1-ATPase subunit ϵ’s role in latent ATPase hydrolysis
Authors: Wong, Chui-Fann
Lau, Aik-Meng
Harikishore, Amaravadhi
Saw, Wuan-Geok
Shin, Joon
Ragunathan, Priya
Bhushan, Shashi
Ngan, So-Fong Cam
Sze, Siu Kwan
Bates, Roderick Wayland
Dick, Thomas
Grüber, Gerhard
Keywords: Science::Biological sciences::Biochemistry
Issue Date: 2021
Source: Wong, C., Lau, A., Harikishore, A., Saw, W., Shin, J., Ragunathan, P., Bhushan, S., Ngan, S. C., Sze, S. K., Bates, R. W., Dick, T. & Grüber, G. (2021). A systematic assessment of mycobacterial F1-ATPase subunit ϵ’s role in latent ATPase hydrolysis. FEBS Journal, 288(3), 818-836. https://dx.doi.org/10.1111/febs.15440
Journal: FEBS Journal
Abstract: In contrast to most bacteria, the mycobacterial F1FO-ATP synthase (α3:β3:γ:δ:ϵ:a:b:b’:c9) does not perform ATP hydrolysis-driven proton translocation. Although subunits α, γ and ϵ of the catalytic F1-ATPase component α3:β3:γ:ϵ have all been implicated in the suppression of the enzyme’s ATPase activity, the mechanism remains poorly defined. Here, we brought the central stalk subunit ϵ into focus by generating the recombinant Mycobacterium smegmatis F1-ATPase (MsF1-ATPase), whose 3D low-resolution structure is presented, and its ϵ-free form MsF1αβγ, which showed an eightfold ATP hydrolysis increase and provided a defined system to systematically study the segments of mycobacterial ϵ’s suppression of ATPase activity. Deletion of four amino acids at ϵ’s N terminus, mutant MsF1αβγϵΔ2-5, revealed similar ATP hydrolysis as MsF1αβγ. Together with biochemical and NMR solution studies of a single, double, triple and quadruple N-terminal ϵ-mutants, the importance of the first N-terminal residues of mycobacterial ϵ in structure stability and latency is described. Engineering ϵ’s C-terminal mutant MsF1αβγϵΔ121 and MsF1αβγϵΔ103-121 with deletion of the C-terminal residue D121 and the two C-terminal ɑ-helices, respectively, revealed the requirement of the very C terminus for communication with the catalytic α3β3-headpiece and its function in ATP hydrolysis inhibition. Finally we applied the tools developed during the study for an in silico screen to identify a novel subunit ϵ-targeting F-ATP synthase inhibitor.
URI: https://hdl.handle.net/10356/149231
ISSN: 1742-4658
DOI: 10.1111/febs.15440
Rights: © 2021 Federation of European Biochemical Societies. All rights reserved. This is the accepted version of the following article: Wong, C., Lau, A., Harikishore, A., Saw, W., Shin, J., Ragunathan, P., Bhushan, S., Ngan, S. C., Sze, S. K., Bates, R. W., Dick, T. & Grüber, G. (2020). A systematic assessment of mycobacterial F1-ATPase subunit ϵ’s role in latent ATPase hydrolysis. FEBS Journal, 288(3), 818-836. https://dx.doi.org/10.1111/febs.15440, which has been published in final form at https://doi.org/10.1111/febs.15440
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Journal Articles

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