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ATP synthase produces ATP, a major energy source for metabolic processes in organisms, from ADP and inorganic phosphate in cellular membranes. ATP synthase is known as a rotary motor, in which the c-subunit ring functions as a rotor. In this work, we have tried to develop a more general preparation procedure of thermophilic Foc-ring (TFoc-ring) for NMR measurements. The expression of TFoF1 is easily affected by various experimental conditions such as temperature, shape and size of a flask, a volume of medium, and shaking rate of an incubator. Accordingly, we have tried to optimize the expression conditions of TFoF1. TFoc-rings were purified from TFoF1 according to a reported method. We modified purification procedures to improve purity and yield of TFoc. On top of them, we found a new combination of detergents for the purification at anion-exchange column chromatography. To examine the effect of lipid environments on the structure, the TFoc-rings were reconstituted into two kinds of lipid bilayers, namely, saturated and unsaturated lipid ones. Then, we have compared characteristics of the TFoc-ring structures in these membranes with solid-state NMR.

journal of the korean magnetic resonance society

Improved Purification of Thermophilic F<sub>o</sub>F<sub>1</sub>-ATP Synthase c-Subunit Rings and Solid-State NMR Characterization of Them in Different Lipid Membranes

Improved Purification of Thermophilic FoF1-ATP Synthase c-Subunit Rings and Solid-State NMR Characterization of Them in Different Lipid Membranes

Improved Purification of Thermophilic F_oF₁-ATP Synthase c-Subunit Rings and Solid-State NMR Characterization of Them in Different Lipid Membranes