ATP Synthesis by Purified ATP-Synthase from Beef Heart Mitochondria After Coreconstitution with Bacteriorhodopsin

Matuschka, Simone; Zwicker, Klaus; Nawroth, Thomas; Zimmer, G

doi:10.1006/abbi.1995.1445

Cited by 16 publications

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“…Spinach chloroplast ATP synthase has also been extensively used to develop ATP synthesis systems (see Table ). In contrast, mitochondrial ATPase has only been used in two studies,123a,134 probably due to difficulties in protein purification compared to spinach or bacterial sources.…”

Section: Intracellular Bioactivities In Artificial Cellsmentioning

confidence: 99%

Toward Artificial Cells: Novel Advances in Energy Conversion and Cellular Motility

Jeong

Nguyen

Kim

et al. 2020

Adv Funct Materials

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The demand to discover every single cellular component has been continuously increasing along with the development of biological techniques. The bottom‐up approach to construct a cell‐mimicking system from well‐defined and tunable compositions is accelerating, with the ultimate goal of comprehending a biological cell. From among the available techniques, the artificial cell has been increasingly recognized as one of the most powerful tools for building a cell‐like system from scratch. This review summarizes the development of artificial cells, from a pure giant unilamellar vesicle (GUV) to a controllable, self‐fueled proteoliposome, both of which are highly compartmentalized. The basic components of an artificial cell, as well as the optimal conditions required for successful, reproducible GUV formation and protein reconstitution, are discussed. Most importantly, progress in studying the metabolic reactions in and the motility of a reconstituted artificial cell are the main focus of the review. The ability to perform a complicated reaction cascade in a controllable manner is highlighted as a promising perspective to obtaining an autonomous and movable GUV.

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Section: Intracellular Bioactivities In Artificial Cellsmentioning

confidence: 99%

Toward Artificial Cells: Novel Advances in Energy Conversion and Cellular Motility

Jeong

Nguyen

Kim

et al. 2020

Adv Funct Materials

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“…In primary structures of Fo (18-25 kDa) and in gamma subunit, the only subunit which exhibits an immediately neighbored basic as well as a hydrophobic residue is the b subunit forming a sequence of Lys-Cys-Ile, labeled increasingly by radioactive lipoic acid in presence of ADP. Therefore, previous studies on oligomycin-sensitive -SH groups in partially purified bovine mitochondrial ATP-synthase [19] and probably also in mitochondria from rat heart and liver can be reconciled and explained by these findings with highly purified ATP-synthase [35] connecting a repair during oxidative stress with increase in thiol reactivity. The results clearly indicate that the reactivity of the -SH group in the b-subunit depends on the enzyme substrate and inhibition.…”

Section: Discussionmentioning

confidence: 65%

“…Isolation of ATPsynthase from bovine heart mitochondria and preparation of submitochondrial particles (SMP) was done according to Matuschka et al [35]. The enzyme in 50 mM Tris, 10% glycerol (v/v), 0.6 mM laurylmaltoside, pH 7.5 (buffer A) was stored in liquid nitrogen.…”

Section: Isolation Of Atpsynthasementioning

confidence: 99%

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ADP‐ and oligomycin‐sensitive redox behavior of F₀ b thiol in ATPsynthase depends on neighbored primary structure: Investigations using 14‐C‐labeled alpha lipoic acid

Dünschede¹,

Zwicker

Ackermann

et al. 2003

BioFactors

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Purified ATPsynthase of bovine heart mitochondria has been analyzed for its mobility and reactivity of oligomycin-sensitive sulfhydryl regions in presence of the substrate ADP and oligomycin. Labeling of thiol groups at the hydrophobic F_0 region of the ATPsynthase was increased in the enzyme initially treated with SDS, N-ethylmaleimide and dithiothreitol (modified enzyme). After dialysis or gel permeation the ATPsynthase was treated with [14C] alpha lipoic acid at a molar ratio of 35-85/1 (lipoic acid/ATPsynthase) corresponding to 4-8.6 nmol/mg protein. Under these conditions, ATPase activity of the native enzyme was significantly decreased. After preincubation with ADP, PAGE of the native, [14C] labeled enzyme revealed an increase of radioactivity at a region of 25 kDa deduced to Cys 197 of subunit b. In the modified enzyme the increase in radioactivity was found at 10 kDa. In this context, the sequence Lys-Cys-Ile around Cys 197 of subunit b suggests excessive reactivity of this thiol, as well as ready reversibility by -SH-S-S- interchange. Therefore, previously observed reaction by thiol reagents and antioxidants from outside the mitochondrion can be interpreted with Cys 197 of F0 b. It accounts for sulfhydryl unmasked by binding of ADP at F1.

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“…In 1974, Racker and Stoeckenius experimentally proved the role of the PMF in Mitchells chemiosmosis theory by coreconstitution of bacteriorhodopsin (bRH) and ATP synthase. By variation of different types of ATP synthases and lipid composition, the stability and productivity of the bRH‐ATP synthase coassembly was continuously improved . In vitro assemblies of bRH and ATP synthase from chloroplasts (CF o F 1 ), beef heart mitochondria (MFoF1), and thermophilic Bacillus PS3 (TFoF1) have been reported (please see Table ).…”

Section: Light Energy Conversion To Atpmentioning

confidence: 99%

Artificial Organelles for Energy Regeneration

et al. 2019

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cell's total energy budget. [2] The universal energy currency that is used for these purposes and can be found in all forms of life is adenosine triphosphate (ATP). A vast majority of the cellular energy demand is covered by either converting a large variety of energy-rich substrates into ATP in a process called oxidative phosphorylation, or by converting light (electromagnetic) energy into ATP in a process called photophosphorylation (or photosynthesis).These naturally existing energy conversions are valid in in the context of bottomup synthetic biology as well. In the latter, an artificial cell is envisioned as a compendium of functional modules, each hand-tailored to partially or entirely mimic one of the essential life processes, such as reproduction, growth, motility, etc. [3] Like their natural counterparts, all these reenvisioned synthetic processes are energy demanding, therefore, the deliberate design of synthetic cells should involve suitable energy management strategy, by, for example, continuous regeneration of ATP. Apart from the importance in the context of artificial cells, new energy conversion strategies can be considered as a standalone feature for enzymatic and cell-free biotechnology, wherein bottom-up synthetic biology might also deliver new and sustainable solutions.The notion of synthetic in terms of engineered and/or non-natural can be even expanded to other forms of energy (like electrical energy) and non-natural building blocks. The chemically driven ATP synthesis, as in oxidative phosphorylation, represents a spontaneous process, in which the electrons of a fuel (glucose, NADH) are transferred to an electron acceptor such as oxygen with the concomitant generation of proton gradient, which is afterwards stored again as chemical energy in ATP. In the realm of synthetic biology, other options might also be feasible, for example: can we use electrical energy and directly plug it in to drive biological processes [4,5] or make use of natural electron transfer mechanisms to produce electricity? [6] The Electron Transport Chain-a Natural Toolbox of Functional Parts for the Construction of Artificial OrganellesDuring the process of oxidative phosphorylation, electrons are passed from an electron donor ("fuel") with a more negative One of the critical steps in sustaining life-mimicking processes in synthetic cells is energy, i.e., adenosine triphosphate (ATP) regeneration. Previous studies have shown that the simple addition of ATP or ATP regeneration systems, which do not regenerate ATP directly from ADP and P i , have no or only limited success due to accumulation of ATP hydrolysis products. In general, ATP regeneration can be achieved by converting light or chemical energy into ATP, which may also involve redox transformations of cofactors. The present contribution provides an overview of the existing ATP regeneration strategies and the related nicotinamide adenine dinucleotide (NAD + ) redox cycling, with a focus on compartmentalized systems. Special attention is being paid to those approach...

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ATP Synthesis by Purified ATP-Synthase from Beef Heart Mitochondria After Coreconstitution with Bacteriorhodopsin

Cited by 16 publications

References 0 publications

Toward Artificial Cells: Novel Advances in Energy Conversion and Cellular Motility

Toward Artificial Cells: Novel Advances in Energy Conversion and Cellular Motility

ADP‐ and oligomycin‐sensitive redox behavior of F₀ b thiol in ATPsynthase depends on neighbored primary structure: Investigations using 14‐C‐labeled alpha lipoic acid

Artificial Organelles for Energy Regeneration

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ATP Synthesis by Purified ATP-Synthase from Beef Heart Mitochondria After Coreconstitution with Bacteriorhodopsin

Cited by 16 publications

References 0 publications

Toward Artificial Cells: Novel Advances in Energy Conversion and Cellular Motility

Toward Artificial Cells: Novel Advances in Energy Conversion and Cellular Motility

ADP‐ and oligomycin‐sensitive redox behavior of F0 b thiol in ATPsynthase depends on neighbored primary structure: Investigations using 14‐C‐labeled alpha lipoic acid

Artificial Organelles for Energy Regeneration

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ADP‐ and oligomycin‐sensitive redox behavior of F₀ b thiol in ATPsynthase depends on neighbored primary structure: Investigations using 14‐C‐labeled alpha lipoic acid