METHOD OF STUDY OF MECHANOCHEMICAL CONVERSIONS OF OLIGOMERIC ENZYMES BY ACOUSTIC RESONANCE METHOD

摘要

FIELD: biophysics, biochemistry, enzymology. SUBSTANCE: for oligomeric membrane-bound enzyme F1-F0 ATPase in submitochondrial particles a method of study of conformation conversions in the process of enzymatic catalysis is shown. These particles are necessary for realization of coupling in vector transport of protons and scalar reaction of ATP synthesis in vivo. According to Curie principle about the impossibility of coupling processes that are tensors of different rank the most possible event during these conformation alternations of F1-F0 ATPase (this process is accompanied by change of affinity to substrates-products of reaction) will be process of reciprocal drawing together-removing factors F1 and F0. In confirmation of oscillatory conformation motions of ATPase domains a possibility of total ATP synthesis (in the absence of <EMI ID=0.501 HE=9 WI=9 TI=CHI>-generator) using acoustic wave as sinusoidal external force is shown. For this purpose the standard acoustic stand including generator of signals, enhancer, dynamic loudspeaker and special experimental cuvette (made as a disk) is used. Cuvette has a solution containing submitochondrial particles and substrate ATP for synthetase reaction. Recording of synthesis of ATP induced by acoustic wave is carried out by standard enzymatic method by reduction of NADP using coupled hexokinase and glucose-6-phosphate dehydrogenase reactions. Dependence of ATP synthesis rate from sound frequency shows two maxima (harmonics) at about the range 170 and 320 Hz with corresponding rate values - 10 and 8 nmoles ATP/min/mg of protein of submitochondrial particles. EFFECT: improved method of investigation. 2 dwg