The uniqueness of subunit α of mycobacterial F-ATP synthases: An evolutionary variant for niche adaptation

Priya Ragunathan, Hendrik Sielaff, Lavanya Sundararaman, Goran Biuković, Malathy Sony Subramanian Manimekalai, Dhirendra Singh, Subhashri Kundu, Thorsten Wohland, Wayne Frasch, Thomas Dick, Gerhard Grüber

    Research output: Contribution to journalArticle

    8 Citations (Scopus)

    Abstract

    The F1F0-ATP (F-ATP) synthase is essential for growth of Mycobacterium tuberculosis, the causative agent of tuberculosis (TB). In addition to their synthase function most F-ATP synthases possess an ATP-hydrolase activity, which is coupled to proton-pumping activity. However, the mycobacterial enzyme lacks this reverse activity, but the reason for this deficiency is unclear. Here, we report that a Mycobacterium-specific, 36-amino acid long C-terminal domain in the nucleotide-binding subunit α (Mtα) of F-ATP synthase suppresses its ATPase activity and determined the mechanism of suppression. First, we employed vesicles to show that in intact membrane-embedded mycobacterial F-ATP synthases deletion of the C-terminal domain enabled ATPase and proton-pumping activity. We then generated a heterologous F-ATP synthase model system, which demonstrated that transfer of the mycobacterial C-terminal domain to a standard F-ATP synthase α subunit suppresses ATPase activity. Single-molecule rotation assays indicated that the introduction of this Mycobacterium-specific domain decreased the angular velocity of the power-stroke after ATP binding. Solution X-ray scattering data and NMR results revealed the solution shape of Mtα and the 3D structure of the subunit α C-terminal peptide 521PDEHVEALDEDKLAKEAVKV540 of M. tubercolosis (Mtα(521–540)), respectively. Together with cross-linking studies, the solution structural data lead to a model, in which Mtα(521–540) comes in close proximity with subunit γ residues 104–109, whose interaction may influence the rotation of the camshaft-like subunit γ. Finally, we propose that the unique segment Mtα(514–549), which is accessible at the C terminus of mycobacterial subunit α, is a promising drug epitope.

    Original languageEnglish (US)
    Pages (from-to)11262-11279
    Number of pages18
    JournalJournal of Biological Chemistry
    Volume292
    Issue number27
    DOIs
    StatePublished - 2017

    Fingerprint

    Adenosine Triphosphatases
    Mycobacterium
    Protons
    Adenosine Triphosphate
    Camshafts
    Angular velocity
    Hydrolases
    X ray scattering
    Mycobacterium tuberculosis
    F1F0-ATP synthase
    Epitopes
    Assays
    Tuberculosis
    Nucleotides
    Stroke
    Nuclear magnetic resonance
    X-Rays
    Membranes
    Amino Acids
    Molecules

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Cell Biology

    Cite this

    Ragunathan, P., Sielaff, H., Sundararaman, L., Biuković, G., Manimekalai, M. S. S., Singh, D., ... Grüber, G. (2017). The uniqueness of subunit α of mycobacterial F-ATP synthases: An evolutionary variant for niche adaptation. Journal of Biological Chemistry, 292(27), 11262-11279. https://doi.org/10.1074/jbc.M117.784959

    The uniqueness of subunit α of mycobacterial F-ATP synthases : An evolutionary variant for niche adaptation. / Ragunathan, Priya; Sielaff, Hendrik; Sundararaman, Lavanya; Biuković, Goran; Manimekalai, Malathy Sony Subramanian; Singh, Dhirendra; Kundu, Subhashri; Wohland, Thorsten; Frasch, Wayne; Dick, Thomas; Grüber, Gerhard.

    In: Journal of Biological Chemistry, Vol. 292, No. 27, 2017, p. 11262-11279.

    Research output: Contribution to journalArticle

    Ragunathan, P, Sielaff, H, Sundararaman, L, Biuković, G, Manimekalai, MSS, Singh, D, Kundu, S, Wohland, T, Frasch, W, Dick, T & Grüber, G 2017, 'The uniqueness of subunit α of mycobacterial F-ATP synthases: An evolutionary variant for niche adaptation' Journal of Biological Chemistry, vol. 292, no. 27, pp. 11262-11279. https://doi.org/10.1074/jbc.M117.784959
    Ragunathan, Priya ; Sielaff, Hendrik ; Sundararaman, Lavanya ; Biuković, Goran ; Manimekalai, Malathy Sony Subramanian ; Singh, Dhirendra ; Kundu, Subhashri ; Wohland, Thorsten ; Frasch, Wayne ; Dick, Thomas ; Grüber, Gerhard. / The uniqueness of subunit α of mycobacterial F-ATP synthases : An evolutionary variant for niche adaptation. In: Journal of Biological Chemistry. 2017 ; Vol. 292, No. 27. pp. 11262-11279.
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    abstract = "The F1F0-ATP (F-ATP) synthase is essential for growth of Mycobacterium tuberculosis, the causative agent of tuberculosis (TB). In addition to their synthase function most F-ATP synthases possess an ATP-hydrolase activity, which is coupled to proton-pumping activity. However, the mycobacterial enzyme lacks this reverse activity, but the reason for this deficiency is unclear. Here, we report that a Mycobacterium-specific, 36-amino acid long C-terminal domain in the nucleotide-binding subunit α (Mtα) of F-ATP synthase suppresses its ATPase activity and determined the mechanism of suppression. First, we employed vesicles to show that in intact membrane-embedded mycobacterial F-ATP synthases deletion of the C-terminal domain enabled ATPase and proton-pumping activity. We then generated a heterologous F-ATP synthase model system, which demonstrated that transfer of the mycobacterial C-terminal domain to a standard F-ATP synthase α subunit suppresses ATPase activity. Single-molecule rotation assays indicated that the introduction of this Mycobacterium-specific domain decreased the angular velocity of the power-stroke after ATP binding. Solution X-ray scattering data and NMR results revealed the solution shape of Mtα and the 3D structure of the subunit α C-terminal peptide 521PDEHVEALDEDKLAKEAVKV540 of M. tubercolosis (Mtα(521–540)), respectively. Together with cross-linking studies, the solution structural data lead to a model, in which Mtα(521–540) comes in close proximity with subunit γ residues 104–109, whose interaction may influence the rotation of the camshaft-like subunit γ. Finally, we propose that the unique segment Mtα(514–549), which is accessible at the C terminus of mycobacterial subunit α, is a promising drug epitope.",
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    AU - Biuković, Goran

    AU - Manimekalai, Malathy Sony Subramanian

    AU - Singh, Dhirendra

    AU - Kundu, Subhashri

    AU - Wohland, Thorsten

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    AU - Dick, Thomas

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