Structure and function of Mycobacterium-specific components of F-ATP synthase subunits α and ε

Bogdanović, Nebojša; Sundararaman, Lavanya; Kamariah, Neelagandan; Tyagi, Anu; Bhushan, Shashi; Ragunathan, Priya; Shin, Joon; Dick, Thomas; Grüber, Gerhard

doi:10.1016/j.jsb.2018.10.006

Cited by 13 publications

(26 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, the solution Nuclear Magnetic Resonance (NMR) structure of the Mtb ε-subunit was resolved [56]. This structure, together with genetic and biochemical studies, showed that the Mtb ε-subunit plays a novel role in ATP synthesis by linking c-ring rotation to ATP synthesis at the α 3 β 3 -headpiece [56][57][58]. Critical to this function is the Mtb ε-subunit's inter-domain amino acid interaction network, which transmits information on c-ring rotation throughout the subunit and to its C-terminus.…”

Section: Disrupting the Mycobacterial F-atp Synthase ε-Subunit's Funcmentioning

confidence: 99%

Re-Understanding the Mechanisms of Action of the Anti-Mycobacterial Drug Bedaquiline

2019

Self Cite

View full text Add to dashboard Cite

Bedaquiline (BDQ) inhibits ATP generation in Mycobacterium tuberculosis by interfering with the F-ATP synthase activity. Two mechanisms of action of BDQ are broadly accepted. A direct mechanism involves BDQ binding to the enzyme's c-ring to block its rotation, thus inhibiting ATP synthesis in the enzyme's catalytic α 3 β 3 -headpiece. An indirect mechanism involves BDQ uncoupling electron transport in the electron transport chain from ATP synthesis at the F-ATP synthase. In a recently uncovered second direct mechanism, BDQ binds to the enzyme's ε-subunit to disrupt its ability to link c-ring rotation to ATP synthesis at the α 3 β 3 -headpiece. However, this mechanism is controversial as the drug's binding affinity for the isolated ε-subunit protein is moderate and spontaneous resistance mutants in the ε-subunit cannot be isolated. Recently, the new, structurally distinct BDQ analogue TBAJ-876 was utilized as a chemical probe to revisit BDQ's mechanisms of action. In this review, we first summarize discoveries on BDQ's mechanisms of action and then describe the new insights derived from the studies of TBAJ-876. The TBAJ-876 investigations confirm the c-ring as a target, while also supporting a functional role for targeting the ε-subunit. Surprisingly, the new findings suggest that the uncoupler mechanism does not play a key role in BDQ's anti-mycobacterial activity.Concentration (MIC) of 0.002-0.013 µg/mL [10,11]). This potent activity holds true in vivo where BDQ has demonstrated accelerated sterilizing activity [12], with four months of BDQ treatment being as efficacious as six months of first-line drug treatment of Mtb-infected mice [13]. Importantly, BDQ also displays bactericidal activity against non-replicating Mtb at therapeutically attainable concentrations [14,15]. In the clinical setting, the usage of BDQ has produced promising results. Studies have shown that the usage of the drug for drug-resistant TB treatment improved sputum conversion and reduced chemotherapy duration and relapse [16][17][18][19]. Hence, new BDQ-containing drug regimens are currently being explored in Phase III clinical trials, such as the STREAM and SimpliciTB trials [20]. One such trial, the Nix-TB trial, resulted in the recent US FDA approval of the BDQ-pretomanid-linezolid six months, all-oral regimen for the treatment of drug-resistant TB [21,22].

show abstract

Section: Disrupting the Mycobacterial F-atp Synthase ε-Subunit's Funcmentioning

confidence: 99%

Re-Understanding the Mechanisms of Action of the Anti-Mycobacterial Drug Bedaquiline

2019

Self Cite

View full text Add to dashboard Cite

show abstract

“…In the present study, the synergistic effect of mycobacteria-specific epitopes (MtαCTD and Mtε) on inhibition of ATPase activity was revealed via the hybrid complex α chi 3 :β 3 :γ:Mtε used as a model system. The complex α chi 3 :β 3 :γ:Mtε was visualized for the first time, using the negative-stain electron microscopy, revealing the binding of the heterologous Mtε as well as the position of the Mtα-specific Cterminal epitope within the 3D reconstructed map (4). The interaction of these mycobacterial epitopes was described using mutagenesis and cysteine crosslinking experiments.…”

Section: List Ofmentioning

confidence: 99%

“…The mutations in Mtε were designed to dissect the role of its C-terminal domain, hinge domain and N-terminal domain in the catalysis and stability of the reconstituted α chi 3 :β 3 :γ:Mtε (x) complexes. The mutations MtεE87A, MtεR62L, and Mtε 1-120 described the coupling and the entrapment of the inhibitory Mg-ADP, while the mutant Mtε 6-121 emphasized the role of MtεNTD for the protein stability in comparison to the other bacterial counterparts (4,5).…”

Section: List Ofmentioning

confidence: 99%

“…3 :β 3 :γ:Mtε during the ATP hydrolysis and assigning the contribution of Mtε to the catalysis. 4. Defining the specific residues and domains of Mtε that affect the catalysis and/or association of the Mtε with the α chi 3 :β 3 :γ 5.…”

Section: Describing the Kinetic Properties Of The α Chimentioning

confidence: 99%

“…The important residues are labeled with one-letter amino acid codes and inter-proton distances less than 5 Å are marked as dashed lines. Figure adapted from (205).…”

Section: Purification Of and The Characterization Of The Mutant Mtεr62lmentioning

confidence: 99%

See 2 more Smart Citations

Characterization of regulatory central stalk epitopes of mycobacterial- and acetogen F-ATP synthases

Bogdanović¹

View full text Add to dashboard Cite

show abstract

C‐terminal regulatory domain of the ε subunit of F_oF₁ ATP synthase enhances the ATP‐dependent H⁺ pumping that is involved in the maintenance of cellular membrane potential in Bacillus subtilis

et al. 2019

View full text Add to dashboard Cite

The ε subunit of FoF1‐ATPase/synthase (FoF1) plays a crucial role in regulating FoF1 activity. To understand the physiological significance of the ε subunit‐mediated regulation of FoF1 in Bacillus subtilis, we constructed and characterized a mutant harboring a deletion in the C‐terminal regulatory domain of the ε subunit (ε∆C). Analyses using inverted membrane vesicles revealed that the ε∆C mutation decreased ATPase activity and the ATP‐dependent H+‐pumping activity of FoF1. To enhance the effects of ε∆C mutation, this mutation was introduced into a ∆rrn8 strain harboring only two of the 10 rrn (rRNA) operons (∆rrn8 ε∆C mutant strain). Interestingly, growth of the ∆rrn8 ε∆C mutant stalled at late‐exponential phase. During the stalled growth phase, the membrane potential of the ∆rrn8 ε∆C mutant cells was significantly reduced, which led to a decrease in the cellular level of 70S ribosomes. The growth stalling was suppressed by adding glucose into the culture medium. Our findings suggest that the C‐terminal region of the ε subunit is important for alleviating the temporal reduction in the membrane potential, by enhancing the ATP‐dependent H+‐pumping activity of FoF1.

show abstract

Structure and function of Mycobacterium-specific components of F-ATP synthase subunits α and ε

Cited by 13 publications

References 48 publications

Re-Understanding the Mechanisms of Action of the Anti-Mycobacterial Drug Bedaquiline

Re-Understanding the Mechanisms of Action of the Anti-Mycobacterial Drug Bedaquiline

Characterization of regulatory central stalk epitopes of mycobacterial- and acetogen F-ATP synthases

C‐terminal regulatory domain of the ε subunit of F_oF₁ ATP synthase enhances the ATP‐dependent H⁺ pumping that is involved in the maintenance of cellular membrane potential in Bacillus subtilis

Contact Info

Product

Resources

About

Structure and function of Mycobacterium-specific components of F-ATP synthase subunits α and ε

Cited by 13 publications

References 48 publications

Re-Understanding the Mechanisms of Action of the Anti-Mycobacterial Drug Bedaquiline

Re-Understanding the Mechanisms of Action of the Anti-Mycobacterial Drug Bedaquiline

Characterization of regulatory central stalk epitopes of mycobacterial- and acetogen F-ATP synthases

C‐terminal regulatory domain of the ε subunit of FoF1 ATP synthase enhances the ATP‐dependent H+ pumping that is involved in the maintenance of cellular membrane potential in Bacillus subtilis

Contact Info

Product

Resources

About

C‐terminal regulatory domain of the ε subunit of F_oF₁ ATP synthase enhances the ATP‐dependent H⁺ pumping that is involved in the maintenance of cellular membrane potential in Bacillus subtilis