The chemical properties of out-of-equilibrium states of proteins and the role of these states in protein functioning.

The out-of-equilibrium states of several iron-containing proteins (cytochromes c of different origin, haemoglobin, myoglobin, ferredoxin and other non-haem iron proteins, cytochrome c oxidase, horseradish peroxidase) were recorded after fast changes in the active centre (electron reduction of iron, ligand dissociation). Strained states result in which the active centre has already been changed and undergone vibrational relaxation but the main part of protein globule is in the 'old', now out-of-equilibrium, state. Protein structure and chemical properties in these states differ considerably from those in equilibrium states. As a rule, the rate constants of protein-specific chemical reactions increase in out-of-equilibrium states by 1--3 orders of magnitude in comparison with those in equilibrium states. Spectra and reactivity of these proteins change in the course of slow (up to 10(-1) s) conformational relaxation, continuously approaching the equilibrium values. It seems that this conformational relaxation is essentially the elementary act of many enzymic reactions for which the rate of substrate-product transformation is determined by the rate of this conformational change.