Fractionation of avian erythrocyte histones by hydrophobic interaction chromatography

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Abstract
The interactions of [goose] H[histone]1 (H1A, H1B), H2A, H2B, H3, H4 and H5 with phenyl cross-linked agarose were studied. Procedures were described whereby all 6 histones could be bound, released and fractionated by using appropriate salt concentrations or pH. The binding could be totally abolished by inclusion of hydrophobic disrupting agents. Control experiments with nonderivated cross-linked agarose ruled out a passive aggregation-disaggregation phenomenon governing the binding patterns. The absorption sequence based on the identification and quantitation of individual histones from unfractionated (whole) histone or separate histone classes was as follows: H3 .gtoreq. H4 > H2B .gtoreq. H5 .gtoreq. H2A > H1A .gtoreq. H1B. The order differed only slightly from the reverse of the desorption sequence. H1B .ltoreq. H1A .ltoreq. H5 < H2A .ltoreq. H2B < H4 .ltoreq. H3. Preferential interaction of H2A-H2B, H3-H4 and H2A-H2B-H4 occur. These interactions can modify the original relative affinity of each individual component for the matrix. The variability in matrix affinity appeared to involve simple stoichiometry of the histone components.