The Effects of anatpERibosome-Binding Site Mutation on the Stoichiometry of the c Subunit in the F1F0ATPase ofEscherichia coli

Schemidt, Randy A.; Hsu, Debbie K.W.; Deckers-Hebestreit, Gabriele; Altendorf, Karlheinz; Brusilow, William S.A.

doi:10.1006/abbi.1995.0063

Cited by 19 publications

(20 citation statements)

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“…In addition, this study addresses the important question of whether organisms occasionally may alter their c-ring stoichiometry in the living cell in response to environmental conditions such as variations in the available carbon source (43,44). Genetically engineered E. coli c-rings with 10 and 12 hairpins indeed yielded active ATP synthases (45,46); however, only marginal ATP synthesis rates were found in the thermophilic bacterium Bacillus PS3 when the stoichiometry of its rotor ring deviated from the native c 10 (47).…”

Section: Discussionmentioning

confidence: 99%

Engineering rotor ring stoichiometries in the ATP synthase

Pogoryelov¹,

Klyszejko

Krasnoselska³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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ATP synthase membrane rotors consist of a ring of c-subunits whose stoichiometry is constant for a given species but variable across different ones. We investigated the importance of c/c-subunit contacts by site-directed mutagenesis of a conserved stretch of glycines (GxGxGxGxG) in a bacterial c 11 ring. Structural and biochemical studies show a direct, specific influence on the c-subunit stoichiometry, revealing c <11 , c 12 , c 13 , c 14 , and c >14 rings. Molecular dynamics simulations rationalize this effect in terms of the energetics and geometry of the c-subunit interfaces. Quantitative data from a spectroscopic interaction study demonstrate that the complex assembly is independent of the c-ring size. Real-time ATP synthesis experiments in proteoliposomes show the mutant enzyme, harboring the larger c 12 instead of c 11 , is functional at lower ion motive force. The high degree of compliance in the architecture of the ATP synthase rotor offers a rationale for the natural diversity of c-ring stoichiometries, which likely reflect adaptations to specific bioenergetic demands. These results provide the basis for bioengineering ATP synthases with customized ion-to-ATP ratios, by sequence modifications.alpha helix packing | F 1 F o ATP synthase | membrane protein | rotary motor stoichiometry | bioenergetics

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Section: Discussionmentioning

confidence: 99%

Engineering rotor ring stoichiometries in the ATP synthase

Pogoryelov¹,

Klyszejko

Krasnoselska³

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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“…Interestingly, there have been reports that the proton stoichiometry in ATPase may vary depending on environmental conditions in Escherichia coli (Schemidt et al, 1995(Schemidt et al, , 1998 The Cyt b 6 /f complex has a unique role in chloroplast electron transport, as it can act in both linear electron transport (production of ATP and NADPH) and cyclic electron transport (ATP generation only). There was a strong linear relationship between chloroplast ETR and Cyt b 6 /f content such that electron transport per Cyt b 6 /f content was the same for plants with a large range of Rieske FeS content (Figs.…”

Section: Atp Synthase Activity Varies In Vivomentioning

confidence: 99%

The Roles of ATP Synthase and the Cytochrome b 6/f Complexes in Limiting Chloroplast Electron Transport and Determining Photosynthetic Capacity

et al. 2010

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In C 3 plants, CO 2 assimilation is limited by ribulose 1,5-bisphosphate (RuBP) regeneration rate at high CO 2 . RuBP regeneration rate in turn is determined by either the chloroplast electron transport capacity to generate NADPH and ATP or the activity of Calvin cycle enzymes involved in regeneration of RuBP. Here, transgenic tobacco (Nicotiana tabacum 'W38') expressing an antisense gene directed at the transcript of either the Rieske iron-sulfur protein of the cytochrome (Cyt) b 6 /f complex or the d-subunit of chloroplast ATP synthase have been used to investigate the effect of a reduction of these complexes on chloroplast electron transport rate (ETR). Reductions in d-subunit of ATP synthase content did not alter chlorophyll, Cyt b 6 /f complex, or Rubisco content, but reduced ETR estimated either from measurements of chlorophyll fluorescence or CO 2 assimilation rates at high CO 2 . Plants with low ATP synthase content exhibited higher nonphotochemical quenching and achieved higher ETR per ATP synthase than the wild type. The proportional increase in ETR per ATP synthase complex was greatest at 35°C, showing that the ATP synthase activity can vary in vivo. In comparison, there was no difference in the ETR per Cyt b 6 /f complex in plants with reduced Cyt b 6 /f content and the wild type. The ETR decreased more drastically with reductions in Cyt b 6 /f complex than ATP synthase content. This suggests that chloroplast ETR is more limited by Cyt b 6 /f than ATP synthase content and is a potential target for enhancing photosynthetic capacity in crops.Plants capture light energy with their light-harvesting systems, including chlorophylls (Chls) and carotenoids, and drive photosynthetic electron transport through the thylakoid membranes of the chloroplasts. Electrons excised from water in PSII are ultimately transferred to NADP + via PSI, resulting in production of NADPH. This process is known as linear electron transport. At the same time, this linear electron transport that passes through the cytochrome (Cyt) b 6 /f complex generates a proton gradient across the thylakoid membrane (DpH;Allen, 2003). Together with the proton gradient generated by the water-splitting complex associated with PSII, these proton gradients enable ATP production by the ATP synthase complex and help to regulate nonphotochemical quenching (NPQ) of excitation energy (Mü ller et al., 2001). There is also a cyclic electron transport that depends on the PSI photochemical reactions and also passes through the Cyt b 6 /f complex. The cyclic electron transport can generate a DpH and drives ATP synthesis by ATP synthase without concomitant generation of NADPH (Shikanai, 2007). ATP and NADPH generated by light reactions are utilized primarily in the Calvin cycle and photorespiratory cycle. The activity and regulation of the Cyt b 6 /f complex and the ATP synthase are thus key components determining the rate of NADPH and ATP production for CO 2 fixation. Photosynthetic CO 2 assimilation rate can be viewed as being limited either by the capacity o...

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“…In the latter model, the c‐subunits each consist of two parallel helices, each set of two connected through a loop. The helices have an annular arrangement of 12 units, but in other organisms higher and lower numbers have been observed as well (see for instance Refs 22–25…”

Section: Introductionmentioning

confidence: 98%

A computational model of the inhibition of Mycobacterium tuberculosis ATPase by a new drug candidate R207910

et al. 2007

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Diarylquinolines (DARQs) are a new class of potent inhibitors of the ATPase of Mycobacterium tuberculosis. We have created a homology model of a binding site for this class of compounds located on the contact area of the a-subunit (gene atpB) and c-subunits (gene atpE) of Mycobacterium tuberculosis ATPase. The binding pocket that was identified from the analysis of the homology model is formed by 4 helices of three c-subunits and 2 helices of the a-subunit. The lead compound of the DARQ series, R207910, was docked into the pocket using a simulated annealing, multiple conformer, docking algorithm. Different stereoisomers were treated separately. The best docking pose for each stereoisomer was optimized by molecular dynamics simulation on the 5300 atoms of the binding region and ligand. The interaction energies in the computed complexes enable us to rank the different stereoisomers in order of interaction strength with the ATPase binding pockets. We propose that the activity of R207910 against Mycobacterium tuberculosis is based on interference of the compound with the escapement geometry of the proton transfer chain. Upon binding the compound mimics the conserved Arg-186 residue of the a-subunit and interacts in its place with the conserved acidic residue Glu-61 of the c-subunit. This mode of action is corroborated by the good agreement between the computed interaction energies and the observed pattern of stereo-specificity in the model of the binding region.

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The Effects of anatpERibosome-Binding Site Mutation on the Stoichiometry of the c Subunit in the F1F0ATPase ofEscherichia coli

Cited by 19 publications

References 0 publications

Engineering rotor ring stoichiometries in the ATP synthase

Engineering rotor ring stoichiometries in the ATP synthase

The Roles of ATP Synthase and the Cytochrome b 6/f Complexes in Limiting Chloroplast Electron Transport and Determining Photosynthetic Capacity

A computational model of the inhibition of Mycobacterium tuberculosis ATPase by a new drug candidate R207910

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