Action of heparin on mammalian nuclei. I. Differential extraction of histone H1 and cooperative removal of histones from chromatin.
Heparin interacts strongly with the histone component of chromatin, forming heparin-histone complexes which resist dissociation by 0.2 M H2SO4. Heparin treatment of unfractionated histones isolated from nuclei of Chinese hamster cells indicates that the affinities of the histone classes for heparin appear in the order from greatest to least: (H3, H4) greater than (H2A, H2B) greater than H1. However, when isolated nuclei are treated with heparin, H1 is released from the chromatin more readily than the other four histone classes. The release of these four histones (H2A, H2B, H3, and H4) is coordinate and occurs in a highly cooperative manner, as indicated by (1) dependence of the initial kinetics of histone removal upon heparin concentration, (2) analysis of DNA and histones in the fractions obtained from differential sedimentation of heparin-treated nuclei, and (3) analysis of the products from heparin-treated nuclei by equilibrium centrifugation in metrizamide density gradients. The results suggest rapid procedures for using heparin as an agent for studying the accessibility of histones in chromatin of intact nuclei. The relationship of these results to current models of chromatin structure is discussed.
D001665:Binding Sites; D002458:Cell Fractionation; D002460:Cell Line; D002467:Cell Nucleus; D002499:Centrifugation, Density Gradient; D002843:Chromatin / Q000032:analysis; D004247:DNA / Q000302:isolation & purification; D006493:Heparin; D006657:Histones / Q000302:isolation & purification; D007700:Kinetics; D011485:Protein Binding; D013696:Temperature
D002843:Chromatin; D006657:Histones; D006493:Heparin; D004247:DNA