[Interconnection between activity and conformational mobility of alpha-chymotrypsin in reverse micelle systems].

A comparative study of the catalytic activity of alpha-chymotrypsin and the spin label rotation frequency in the alpha-chymotrypsin active center of reverse micellar systems solvated by H2O-organic mixtures was carried out. It was found that the decrease in the label rotation frequency resulting from the substitution of water in the micellar inner cavity by glycerol, 2.3-butanediol and dimethylsulfoxide (up to 95%) caused a marked increase (20-fold in the case of 2.3-butanediol) of the enzyme catalytic activity. The experimental results are quantitatively interpreted in terms of a simple kinetic scheme postulating the existence of the enzyme in two interconvertible forms differing in the conformational (intramolecular) mobility, i.e., the resting one predominantly existing in aqueous solution, and the tense one whose proportion rises with an increase in the concentration of the water-miscible organic solvent in the reverse micellar system. The value of kcat (2.4 s-1) for the tense form of the enzyme exceeded by more than 25 times the catalytic activity of chymotrypsin in aqueous solution (0.09 s-1) for the resting form.