Methodical aspects of determining the oxidative modification of the protein

Abstract

Introduction. To date, a large amount of data has been accumulated on oxidative stress, its inducers and its effect on the human body. In this article, the main changes in protein molecules under the influence of products of oxidative stress are reviewed, and a brief review of the main methods for detecting protein’s modification was made. Aim of investigation - selection of some methodical approaches to determination of level of oxidative modification of protein of blood serum. Materials and methods. A modification of the Reznick et al. method to determine the oxidative modification of the protein in a microversion, the essence of which is a proportional decrease in the volumes of reagents participating in the reaction and in the replacement of certain reagents with their analogues in chemical nature. Results. The concentration of serum ketone-dinitrophenylhydrazones of neutral type (kDNPn) of healthy peripheral blood serum was 1.24±0.15, the ketone-dinitrophenylhydrazones of basic type (kDNPb) were 0.74±0.11, and the aldehyde-dinitrophenylhydrazones of neutral type (aDNPn) 0.37±0.07 nmol/mg protein. The total total concentration of oxidized proteins of peripheral blood serum of healthy donors was 2.35±0.11 nmol/mg protein. It was revealed that aDNPn amount for 16%, for kDNPb - 31.5%, for kDNPn - 52.5% of oxidized proteins. Thus, the ratio of oxidized proteins of blood serum of healthy donors to aDNPn : kDNPb : kDNPn is 1 : 2 : 3.3. Conclusion. During the research it was found that when the level of oxidized protein is determined by a micro-method, the level of oxidized proteins corresponds to the results obtained both in the classical variant of the reaction and in the literature data.