Tag Archives: equilibrium constant
Consideration of the contribution of chemical (non-enzymatic) conversion of substrate in the general mechanism of enzyme reaction
S. O. Kosterin, S. O. Karakhim, P. F. Zhuk
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: kinet@biochem.kiev.ua; laserlab@biochem.kiev.ua
Received: 13 September 2018; Accepted: 13 December 2018
When enzyme-catalyzed reactions are studied, it is necessary to take into account the contribution of the chemical (non-enzymatic) conversion of the substrate to the product, which is carried out together with the enzyme-catalyzed conversion of the substrate. It is generally believed that the difference of the product concentration that was formed in the presence of the enzyme and in its absence (during the same time interval) is the concentration of the product that was formed directly in the enzyme-catalyzed reaction, i.e. that there is additivity of the product concentrations at each time point. In this paper, we have analyzed when there is additivity and how to correctly take into account the contribution of chemical (non-catalytic) substrate conversion when the enzyme-catalyzed reactions are investigated. We have shown that the additivity of product concentrations and initial rates is observed only for a period when the product concentration increases linearly with time. The longer the reaction proceeds the more the deviation from the additivity. Under equilibrium condition, there is no additivity of equilibrium product concentrations but under conditions of detailed balance the equilibrium product concentration of the overall reaction, including the enzyme-catalyzed and chemical (non-enzymatic) conversion of the substrate, is also at the same time the equilibrium concentration of the product of the enzyme-catalyzed conversion of the substrate.
New approach in evaluating affinity of bivalent antibodies by the method of surface plasmon resonance. Theory
S. A. Bobrovnik, M. O. Demchenko, S. . Komisarenko
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv;
e-mail: s-bobrov@bk.ru
Theoretical aspects of the affinity evaluation for the interaction between bivalent receptors (or antibodies) and corresponding ligands (or antigens) are considered. It was shown that the ligand presence in the solution at the stage when the receptor dissociation occurs leads to the increase of the affinity evaluation accuracy. We demonstrated that the analysis of the dissociative curve of the receptor from the chip is not necessary for affinity determination; the analysis of associative curve is sufficient for this purpose. We also suggested a new approach for evaluating the affinity of bivalent receptors (or antibodies) when these reagents are present in the studied solution and the correspondent ligand (or antigen) is immobilized on the chip.