scholarly journals Functional characterization of mutants deficient in N-terminal phosphorylation of the CF1 subunit beta

2021 ◽  
Author(s):  
Ralph Bock ◽  
Deserah D Strand ◽  
Daniel Karcher ◽  
Stephanie Ruf ◽  
Anne Schadach ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Electron Transport ◽  
Atp Synthase ◽  
Electron Transport Chain ◽  
Atp Hydrolysis ◽  
Molecular Motor ◽  
Functional Characterization ◽  
Phosphorylation Sites ◽  
Transport Chain

Understanding the regulation of photosynthetic light harvesting and electron transfer is of great importance to efforts to improve the ability of the electron transport chain to supply downstream metabolism. The central regulator of the electron transport chain is the ATP synthase, the molecular motor that harnesses the chemiosmotic potential generated from proton coupled electron transport to synthesize ATP. The ATP synthase is regulated both thermodynamically and post-translationally, with proposed phosphorylation sites on multiple subunits. In this study we focused on two N-terminal serines on the catalytic subunit beta, previously proposed to be important for dark inactivation of the complex to avoid ATP hydrolysis at night. Here we show that there is no clear role for phosphorylation in the dark inactivation of ATP synthase. Instead, mutation of one of the two phosphorylated serine residues to aspartate strongly decreased ATP synthase abundance. We propose that the loss of N-terminal phosphorylation of ATP beta may be involved in proper ATP synthase accumulation during complex assembly.

scholarly journals Cytochrome c550 from Pseudomonas aeruginosa

1993 ◽  
Vol 289 (1) ◽  
pp. 173-178 ◽  
Author(s):  
P Reichmann ◽  
H Görisch
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Electron Transfer ◽  
Oxygen Consumption ◽  
Electron Transport ◽  
Electron Transport Chain ◽  
Prosthetic Group ◽  
Midpoint Potential ◽  
Transport Chain ◽  
Ethanol Dehydrogenase ◽  
Soluble C

In cells of Pseudomonas aeruginosa A.T.C.C. 17933 grown on ethanol the synthesis of a soluble c-type cytochrome, together with quinoprotein ethanol dehydrogenase, is induced. The cytochrome, with an alpha-absorption band at 550 nm, was purified to homogeneity. The molecular mass of the monomeric protein is 15 kDa, the pI is 4.8, and it contains one haem prosthetic group. The midpoint potential of the autoxidizable, but not autoreducible, cytochrome is 280 mV. Cytochrome c550 mediates electron transfer between quinoprotein ethanol dehydrogenase and ferricyanide. In a system composed of membrane particles with NN‘NN’-tetramethyl-p-phenylenediamine oxidase activity and quinoprotein ethanol dehydrogenase, oxygen consumption is only observed in the presence of cytochrome c550. This indicates the participation of the cytochrome in the electron-transport chain linked to quinoprotein ethanol dehydrogenase in P. aeruginosa. The electron transport from ethanol dehydrogenase to oxygen is inhibited by myxothiazol and antimycin, indicating that a cytochrome bc1-like complex is involved.

Functional Expression of Electron Transport Chain and FoF1-ATP Synthase in Optic Nerve Myelin Sheath

2015 ◽  
Vol 40 (11) ◽  
pp. 2230-2241 ◽  
Author(s):  
Martina Bartolucci ◽  
Silvia Ravera ◽  
Greta Garbarino ◽  
Paola Ramoino ◽  
Sara Ferrando ◽  
...  
Keyword(s):  
Electron Transport ◽  
Optic Nerve ◽  
Atp Synthase ◽  
Myelin Sheath ◽  
Electron Transport Chain ◽  
Functional Expression ◽  
Transport Chain ◽  
Fof1 Atp Synthase

scholarly journals Experimental conditions affect the site of tetrazolium violet reduction in the electron transport chain of Lactococcus lactis

Microbiology ◽  
2009 ◽  
Vol 155 (9) ◽  
pp. 2941-2948 ◽  
Author(s):  
Sybille Tachon ◽  
Damien Michelon ◽  
Emilie Chambellon ◽  
Monique Cantonnet ◽  
Christine Mezange ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Electron Transport ◽  
Lactococcus Lactis ◽  
Electron Transport Chain ◽  
Experimental Conditions ◽  
Plate Test ◽  
Tetrazolium Salts ◽  
Transport Chain ◽  
Reduction Mechanisms ◽  
Tetrazolium Violet

The reduction of tetrazolium salts to coloured formazans is often used as an indicator of cell metabolism during microbiology studies, although the reduction mechanisms have never clearly been established in bacteria. The objective of the present study was to identify the reduction mechanisms of tetrazolium violet (TV) in Lactococcus lactis using a mutagenesis approach, under two experimental conditions generally applied in microbiology: a plate test with growing cells, and a liquid test with non-growing (resting) cells. The results showed that in both tests, TV reduction resulted from electron transfer from an intracellular donor (mainly NADH) to TV via the electron transport chain (ETC), but the reduction sites in the ETC depended on experimental conditions. Using the plate test, menaquinones were essential for TV reduction and membrane NADH dehydrogenases (NoxA and/or NoxB) were partly involved in electron transfer to menaquinones. In this case, TV reduction mainly occurred outside the cells and in the outer part of the plasma membrane. During the liquid test, TV was directly reduced by NoxA and/or NoxB, probably in the inner part of the membrane, where NoxA and NoxB are localized. In this case, reduction was directly related to the intracellular NADH pool. Based on these findings, new applications for TV tests are proposed, such as NADH pool determination with the liquid test and the screening of mutants affected in menaquinone biosynthesis with the plate test. Preliminary results using other tetrazolium salts in the plate test showed that the reduction sites depended on the salt, suggesting that similar studies should be carried out with other tetrazolium salts so that the outcome of each test can be interpreted correctly.

The efficiency and plasticity of mitochondrial energy transduction

2005 ◽  
Vol 33 (5) ◽  
pp. 897-904 ◽  
Author(s):  
M.D. Brand
Keyword(s):  
Electron Transport ◽  
Atp Synthase ◽  
Electron Transport Chain ◽  
Uncoupling Protein ◽  
Coupling Efficiency ◽  
Energy Transduction ◽  
Proton Pumping ◽  
Complex Iii ◽  
Proton Pumps ◽  
Transport Chain

Since it was first realized that biological energy transduction involves oxygen and ATP, opinions about the amount of ATP made per oxygen consumed have continually evolved. The coupling efficiency is crucial because it constrains mechanistic models of the electron-transport chain and ATP synthase, and underpins the physiology and ecology of how organisms prosper in a thermodynamically hostile environment. Mechanistically, we have a good model of proton pumping by complex III of the electron-transport chain and a reasonable understanding of complex IV and the ATP synthase, but remain ignorant about complex I. Energy transduction is plastic: coupling efficiency can vary. Whether this occurs physiologically by molecular slipping in the proton pumps remains controversial. However, the membrane clearly leaks protons, decreasing the energy funnelled into ATP synthesis. Up to 20% of the basal metabolic rate may be used to drive this basal leak. In addition, UCP1 (uncoupling protein 1) is used in specialized tissues to uncouple oxidative phosphorylation, causing adaptive thermogenesis. Other UCPs can also uncouple, but are tightly regulated; they may function to decrease coupling efficiency and so attenuate mitochondrial radical production. UCPs may also integrate inputs from different fuels in pancreatic β-cells and modulate insulin secretion. They are exciting potential targets for treatment of obesity, cachexia, aging and diabetes.

scholarly journals Tracking Electron Uptake from a Cathode into Shewanella Cells: Implications for Energy Acquisition from Solid-Substrate Electron Donors

mBio ◽  
2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Annette R. Rowe ◽  
Pournami Rajeev ◽  
Abhiney Jain ◽  
Sahand Pirbadian ◽  
Akihiro Okamoto ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Electron Transport ◽  
Complex I ◽  
Electron Transport Chain ◽  
Electron Flow ◽  
Extracellular Electron Transfer ◽  
Terminal Electron Acceptor ◽  
Cellular Energy ◽  
Transport Chain ◽  
Energy Acquisition

ABSTRACTWhile typically investigated as a microorganism capable of extracellular electron transfer to minerals or anodes,Shewanella oneidensisMR-1 can also facilitate electron flow from a cathode to terminal electron acceptors, such as fumarate or oxygen, thereby providing a model system for a process that has significant environmental and technological implications. This work demonstrates that cathodic electrons enter the electron transport chain ofS. oneidensiswhen oxygen is used as the terminal electron acceptor. The effect of electron transport chain inhibitors suggested that a proton gradient is generated during cathode oxidation, consistent with the higher cellular ATP levels measured in cathode-respiring cells than in controls. Cathode oxidation also correlated with an increase in the cellular redox (NADH/FMNH2) pool determined with a bioluminescence assay, a proton uncoupler, and a mutant of proton-pumping NADH oxidase complex I. This work suggested that the generation of NADH/FMNH2under cathodic conditions was linked to reverse electron flow mediated by complex I. A decrease in cathodic electron uptake was observed in various mutant strains, including those lacking the extracellular electron transfer components necessary for anodic-current generation. While no cell growth was observed under these conditions, here we show that cathode oxidation is linked to cellular energy acquisition, resulting in a quantifiable reduction in the cellular decay rate. This work highlights a potential mechanism for cell survival and/or persistence on cathodes, which might extend to environments where growth and division are severely limited.IMPORTANCEThe majority of our knowledge of the physiology of extracellular electron transfer derives from studies of electrons moving to the exterior of the cell. The physiological mechanisms and/or consequences of the reverse processes are largely uncharacterized. This report demonstrates that when coupled to oxygen reduction, electrode oxidation can result in cellular energy acquisition. This respiratory process has potentially important implications for how microorganisms persist in energy-limited environments, such as reduced sediments under changing redox conditions. From an applied perspective, this work has important implications for microbially catalyzed processes on electrodes, particularly with regard to understanding models of cellular conversion of electrons from cathodes to microbially synthesized products.

Localized energy coupling during photophosphorylation by chromatophores of Rhodopseudomonas capsulata N22

1982 ◽  
Vol 2 (10) ◽  
pp. 743-749 ◽  
Author(s):  
G. Duncan Hitchens ◽  
Douglas B. Kell
Keyword(s):  
Free Energy ◽  
Electron Transport ◽  
Atp Synthase ◽  
Electron Transport Chain ◽  
Energy Coupling ◽  
Rhodopseudomonas Capsulata ◽  
Titration Method ◽  
Dual Inhibitor ◽  
Transport Chain ◽  
Synthase Inhibitor

The principle of the dual inhibitor titration method for testing models of electron-transport phosphorylation is outlined, and the method is applied to the study of photophosphorylation in bacterial chromatophores. It is concluded that energy coupling is strictly localized in nature in this system, in the sense that free energy released by a particular electron-transport chain may be used only by a particular H+-ATP synthase. Dual inhibitor titrations using the uncoupler SF 6847 and the H+-ATP synthase inhibitor oligomycin indicate that uncouplers act by shuttling rapidly between the localized energy-coupling sites.

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