Bottom-Up Construction of a Minimal System for Cellular Respiration and Energy Regeneration
Autor: | Judy Hirst, Justin G. Fedor, Zhan Yin, Olivier Biner |
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Rok vydání: | 2020 |
Předmět: |
0106 biological sciences
Alternative oxidase Cellular respiration Biomedical Engineering Respiratory chain 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) Article Mitochondrial Proteins 03 medical and health sciences chemistry.chemical_compound Adenosine Triphosphate ATP hydrolysis 010608 biotechnology Plant Proteins 030304 developmental biology 0303 health sciences Electron Transport Complex I Cell-Free System ATP synthase biology General Medicine Mitochondrial Proton-Translocating ATPases NAD Mitochondria Mitochondrial respiratory chain chemistry Liposomes biology.protein Biophysics Synthetic Biology NAD+ kinase Energy Metabolism Oxidoreductases Adenosine triphosphate |
Zdroj: | ACS Synth Biol |
ISSN: | 2161-5063 |
DOI: | 10.1021/acssynbio.0c00110 |
Popis: | Adenosine triphosphate (ATP), the cellular energy currency, is essential for life. The ability to provide a constant supply of ATP is therefore crucial for the construction of artificial cells in synthetic biology. Here, we describe the bottom-up assembly and characterization of a minimal respiratory system that uses NADH as a fuel to produce ATP from ADP and inorganic phosphate, and is thus capable of sustaining both upstream metabolic processes that rely on NAD(+), and downstream energydemanding processes that are powered by ATP hydrolysis. A detergent-mediated approach was used to co-reconstitute respiratory mitochondrial complex I and an F-type ATP synthase into nanosized liposomes. Addition of the alternative oxidase to the resulting proteoliposomes produced a minimal artificial “organelle” that reproduces the energy-converting catalytic reactions of the mitochondrial respiratory chain: NADH oxidation, ubiquinone cycling, oxygen reduction, proton pumping, and ATP synthesis. As a proof-of-principle, we demonstrate that our nanovesicles are capable of using an NAD(+)-linked substrate to drive cell-free protein expression. Our nanovesicles are both efficient and durable and may be applied to sustain artificial cells in future work. |
Databáze: | OpenAIRE |
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