A5025043446- ATP Synthase and ATPases Research
- Biochemical and Molecular Research
- Tuberculosis Research and Epidemiology
- RNA and protein synthesis mechanisms
- Mitochondrial Function and Pathology
- Mycobacterium research and diagnosis
- RNA modifications and cancer
- Natural Antidiabetic Agents Studies
- Crystallization and Solubility Studies
- X-ray Diffraction in Crystallography
- Cancer therapeutics and mechanisms
- Helicobacter pylori-related gastroenterology studies
- Amoebic Infections and Treatments
- Phytochemicals and Antioxidant Activities
- HIV Research and Treatment
- Photosynthetic Processes and Mechanisms
- Synthesis and biological activity
- Tea Polyphenols and Effects
- Neurological diseases and metabolism
- Enzyme Structure and Function
- Advanced Electron Microscopy Techniques and Applications
- Fibroblast Growth Factor Research
- Genomics and Chromatin Dynamics
- Aquatic life and conservation
- Pharmacological Effects of Natural Compounds
Nanyang Technological University
2012-2024
Abstract Pyrazinamide is a sterilizing first-line tuberculosis drug. Genetic, metabolomic and biophysical analyses previously demonstrated that pyrazinoic acid, the bioactive form of prodrug pyrazinamide (PZA), interrupts biosynthesis coenzyme A in Mycobacterium by binding to aspartate decarboxylase PanD. While most drugs act inhibiting protein function upon target binding, we find here acid only weak enzyme inhibitor. We show PanD triggers degradation caseinolytic protease ClpC1-ClpP. Thus,...
Previously, we showed that a major in vitro and vivo mechanism of resistance to pyrazinoic acid (POA), the bioactive component critical tuberculosis (TB) prodrug pyrazinamide (PZA), involves missense mutations aspartate decarboxylase PanD, an enzyme required for coenzyme A biosynthesis. What is action POA? Upon demonstrating treatment M. bovis BCG with POA resulted depletion intracellular confirming this POA-mediated prevented by either PanD or exogenous supplementation pantothenate,...
The antituberculosis drug bedaquiline (BDQ) inhibits Mycobacterium tuberculosis F-ATP synthase by interfering with two subunits. Drug binding to the c subunit stalls rotation of ring, while ε blocks coupling ring ATP synthesis at catalytic α 3 :β headpiece. BDQ is used for treatment drug-resistant tuberculosis.
The F 1 O ‐ ATP synthase is one of the enzymes that essential to meet energy requirement both proliferating aerobic and hypoxic dormant stages life cycle mycobacteria. Most F‐ synthases consume in α 3 :β headpiece drive γ subunit, which couples cleavage with proton pumping c ring via bottom subunit. ase‐driven H + latent presence a unique 14 amino acid residue loop mycobacterial subunit has been described aligned close vicinity ‐ring Priya R et al . (2013) J Bioenerg Biomembr 45, 121‐129...
The F1 FO -ATP synthase is required for growth and viability of Mycobacterium tuberculosis a validated clinical target. A mycobacterium-specific loop the enzyme's rotary γ subunit plays role in coupling ATP synthesis within enzyme complex. We report discovery novel antimycobacterial, termed GaMF1, that targets this loop. Biochemical NMR studies show GaMF1 inhibits activity by binding to bactericidal active against multidrug- as well bedaquiline-resistant strains. Chemistry efforts on...
Mycobacterium tuberculosis (Mt) F1 F0 ATP synthase (α3 :β3 :γ:δ:ε:a:b:b':c9 ) is essential for the viability of growing and nongrowing persister cells pathogen. Here, we present first NMR solution structure Mtε, revealing an N-terminal β-barrel domain (NTD) a C-terminal (CTD) composed helix-loop-helix with helix 1 -2 being shorter compared to their counterparts in other bacteria. The amino acids are oriented toward NTD, forming domain-domain interface between NTD CTD. Mtε provides novel...
The F
Despite the highly oxidative environment of phagosomal lumen, need for maintaining redox homeostasis is a critical aspect mycobacterial biology. The pathogens are equipped with sophisticated thioredoxin- (Trx) and peroxiredoxin system, including TrxC alkyl hydroperoxide reductase subunit C (AhpC), whereby one reducing partners AhpC. Here we visualize modulated dodecamer ring formation AhpC from Mycobacterium bovis (BCG strain; MbAhpC) using electron microscopy present novel insights into...
The dynamic interaction of the N- and C-terminal domains mycobacterial F-ATP synthase subunit ε is proposed to contribute efficient coupling H+-translocation ATP synthesis. Here, we investigate crosstalk between both by introducing chromosomal atpC missense mutations in helix 2 predicted disrupt inter domain ε-α therefore coupling. mutant εR105A,R111A,R113A,R115A (ε4A) showed decreased intracellular ATP, slower growth rates lower molar yields on non-fermentable carbon sources. Cellular...
The diarylquinoline F 1 O -ATP synthase inhibitor bedaquiline (BDQ) displays protonophore activity. Thus, uncoupling electron transport from ATP synthesis appears to be a second mechanism of action this antimycobacterial drug. Here, we show that the new BDQ analogue TBAJ-876 did not retain parental drug’s Comparative time-kill analyses revealed both compounds exert same bactericidal
The F1FO-ATP synthase engine is essential for viability and growth of non-tuberculous mycobacteria (NTM) by providing the biological energy ATP keeping homeostasis under hypoxic stress conditions. Here, we report discovery diarylquinoline TBAJ-5307 as a broad spectrum anti-NTM inhibitor, targeting FO-domain preventing rotation proton-translocation. active at low nanomolar concentrations against fast- slow-growing NTM well clinical isolates depleting intrabacterial ATP. As demonstrated fast...
The F 1 O -ATP synthase is required for the viability of tuberculosis (TB) and nontuberculous mycobacteria (NTM) has been validated as a drug target. Here, we present cryo-EM structures Mycobacterium smegmatis -ATPase with different nucleotide occupation within catalytic sites visualize critical elements latent ATP hydrolysis efficient synthesis.
While many bacteria are able to bypass the requirement for oxidative phosphorylation when grown on carbohydrates, Mycobacterium tuberculosis is unable do so. Differences of amino acid composition and structural features mycobacterial F-ATP synthase (α3:β3:γ:δ:ε:a:b:b′:c9) compared its prokaryotic or human counterparts were recently elucidated paved avenues discovery molecules interfering with various regulative mechanisms this essential energy converter. In context, peripheral stalk subunit...
In contrast to most bacteria, the mycobacterial F1 FO -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 catalytic -ATPase component α3 :γ:ε have all been implicated in suppression enzyme's ATPase activity, mechanism remains poorly defined. Here, we brought central stalk subunit into focus by generating recombinant Mycobacterium smegmatis (MsF1 -ATPase), whose 3D low-resolution structure is presented, its...
Mycobacterium abscessus (Mab) is a nontuberculous mycobacterium of increasing clinical relevance. The rapidly growing opportunistic pathogen intrinsically multi-drug-resistant and causes difficult-to-cure lung disease. Adenosine triphosphate, generated by the essential F1 FO ATP synthase, major energy currency pathogen, bringing this enzyme complex into focus for discovery novel antimycobacterial compounds. Coupling proton translocation through membrane-embedded sector formation in headpiece...
Subunit b, the peripheral stalk of bacterial F(1)F(o) ATP synthases, is composed a membrane-spanning and soluble part. The part divided into tether, dimerization, delta-binding domains. first solution structure b30-82, including tether region dimerization domain, has been solved by nuclear magnetic resonance, revealing an alpha-helix between residues 39 72. In structure, b30-82 length 48.07 A. surface charge distribution shows one side with hydrophobic pattern, formed alanine residues....
Mycobacteria regulate their energy (ATP) levels to sustain survival even in stringent living conditions. Recent studies have shown that mycobacteria not only slow down respiratory rate but also block ATP hydrolysis of the F-ATP synthase (α3:β3:γ:δ:ε:a:b:b':c9) maintain homeostasis situations amenable for growth. The mycobacteria-specific α C-terminus (α533-545) has unraveled be major regulative latent hydrolysis. Its deletion stimulates ATPase activity while reducing synthesis. In one six...
ABSTRACT The Mycobacterium tuberculosis ( Mtb ) F-ATP synthase generates most of the biological energy currency ATP. Previously, we identified mycobacterium-specific loop subunit γ as a new anti-tuberculosis target and discovered novel diaminopyrimidine GaMF1, whose potency was improved by structure-activity relationship studies leading to analog GaMF1.39. Here, report that GaMF1.39 depletes cellular ATP formation targeting mycobacterial without affecting proton coupling or oxygen...
A common strategy employed in antibacterial drug discovery is the targeting of biosynthetic processes that are essential and specific for pathogen. Specificity particular avoids undesirable interactions with potential enzymatic counterparts human host, it ensures on-target toxicity. Synthesis pantothenate (Vitamine B5), which a precursor acyl carrier coenzyme A, an example such pathway. In Mycobacterium tuberculosis (Mtb), causative agent (TB), formed by synthase, utilizing D-pantoate β-Ala...
New drug targets and molecules with bactericidal activity are needed against the respiratory mycobacterial pathogen Mycobacterium abscessus . Employing a lead repurposing strategy, antituberculosis compound GaMF1 was tested M.
A critical point in the V1 sector and entire V1VO complex is interaction of stalk subunits G (Vma10p) E (Vma4p). Previous work, using precipitation assays, has shown that both form a complex. In this we have analysed N-terminal segment subunit (G1–59) ATPase from Saccharomyces cerevisiae by nuclear magnetic resonance (NMR) spectroscopy. Analyses 1H-15N heteronuclear single quantum coherence (HSQC) spectra G1–59 absence presence peptides E1–18 E18–38 as well produced purified C-terminal...