Gel Chromatography and Gel Electrophoresis of Histones in Denaturing Solvents

Limited Dependence on Molecular Weight

Abstract

Chromatography on polyacrylamide gel (various Bio-Gels) in HCl, guanidine hydrochloride (GuCl), urea, or sodium dodecyl sulfate (SDS) solution and electrophoresis on polyacrylamide gel in acetic acid, acetic acid-urea, or SDS solution were examined for the separation of disulfide bond-reduced histones by molecular weight. Almost complete resolution into five or six molecular species of calf thymus, chicken erythrocyte, or Tetrahymena histone was observed on gel electrophoresis in acidic urea and alkaline SDS solutions, while gel chromatography resulted in poorer resolution. A linear relationship between the known molecular weights of calf histone species and their elution or migration behaviors (distribution or retardation coefficient) was only observed on GuCl-gel chromatography and urea-gel electrophoresis, providing a standard curve for the estimation of unknown molecular weights of chicken and protozoan histone species. On the other gels, certain histone species behaved unexpectedly, especially on gel electrophoresis in acetic acid or SDS solution, in detailed experiments. The reasons for this are discussed.