Histone H1: ultracentrifugation studies of the effects of ionic strength and denaturants on the solution conformation.
The conformation of histone H1 has been examined under native and denaturing conditions in the absence of DNA or chromatin. Sedimentation coefficients were determined for Histone H1 in 0.1 M KCl and in 6 M guanidine hydrochloride solutions at pH 7.4. The influence of ionic strength on the conformation of histone H1 has been determined by measurement of the sedimentation coefficient in tetramethylammonium chloride solutions of up to 2.5 M and extrapolated to infinite ionic strength. Results from these experiments suggest that the native conformation of histone H1 is very asymmetric in shape. The molecule is best described as a prolate ellipsoid with axes of 312 A (2a) and 16 A (2b) in low ionic strength media and also as a prolate ellipsoid with axes of 202 A (2a) and 20 A (2b) at high ionic strength or when associated with polyanions, e.g., DNA. Denaturation of histone H1 by guanidine hydrochloride was found to be completely reversible. In 6 M guanidine hydrochloride, the H1 molecule collapses to a sphere but the original extended conformation of the protein is readily restored on dialysis. This suggests rigid conformational requirements for the H1 molecule as incorporated into chromatin. The shape and dimensions for the H1 molecule at high ionic strength are not sufficiently conclusive to locate H1 in the chromatin structure. It is proposed, however, that viable models for chromatin architecture must be consistent with the histone H1 solution dimensions obtained here.
D006146:Guanidines / Q000494:pharmacology; D006657:Histones; D009994:Osmolar Concentration; D011487:Protein Conformation; D011489:Protein Denaturation; D014461:Ultracentrifugation
D006146:Guanidines; D006657:Histones