Chapter 9 Quantitative Determination of Nonhistone and Histone Proteins in Chromatin

Abstract

Publisher Summary This chapter surveys the most common, existing analytical methods for quantitative nonhistone determinations, and introduces a procedure that is more advantageous in the analysis of the nonhistone moiety of nucleoprotein materials, especially in the presence of histones. The existing methods for quantitative determination of the nonhistone moiety in nucleoprotein material include (1) selective solubilization of histones and precipitation of DNA and nonhistones with dilute mineral acids, (2) quantitative determination of the nonhistones with polyacrylamide gel electrophoresis, and (3) quantitative determination after preparative fractionation of chromatin proteins. The cellogel electrophoresis allows a quantitative determination of chromatin nonhistones in the presence of histones. This method is based mainly on cellogel electrophoresis originally developed for histone analysis. In this method a prerequisite for the analyses is the fractionation of the nucleoprotein complex into the nucleic acids and the protein moiety. The analysis of chromosomal proteins by cellogel electrophoresis after a complete, prior separation of the protein moiety from the DNA by ultracentrifugation in high salt and urea concentrations represents a simple and very reliable way to determine histone and nonhistone quantities. The chapter also provides the examples of the quantitative determination of histones and nonhistones by cellogel electrophoresis.