Chromatin-bound protease: degradation of chromosomal proteins under chromatin dissociation conditions.
A chromatin-bound protease, active in 2 M NaCl-5 M urea or 5 M urea alone, was demonstrated in rat liver, kidney, testes, brain, rabbit bone marrow, chicken reticulocyte, and Ehrlich ascites chromatin. Chicken erythrocyte chromatin did not possess any detectable proteolytic activity in salt and urea. The proteolytic activity of rat liver chromatin in salt and urea was found to be independent of the methods of chromatin preparation. The protease can be inhibited by the serine specific reagents phenylmethanesulfonyl fluoride and diisopropyl fluorophosphate and the alkylating reagent, carbobenzoxyphenylalanine chloromethyl ketone, in the presence of organic solvents at 1 mM concentration. The inhibitions of chromatin-bound protease in rat liver by these compounds are irreversible. On the other hand, carbobenzoxyphenylalanine and p-nitrophenyl acetate were shown to be reversible inhibitors of rat liver chromatin-bound protease. The application of these inhibitors during the dissociation of chromatin by salt and urea may be useful to researchers interested in purifying various chromosomal proteins or to those researchers doing reconstitution studies with labile chromatins.
D000818:Animals; D001853:Bone Marrow / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D002645:Chickens; D002843:Chromatin / Q000201:enzymology*; D002875:Chromosomes / Q000378:metabolism*; D004912:Erythrocytes / Q000378:metabolism; D006657:Histones / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000378:metabolism; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008297:Male; D009928:Organ Specificity; D009994:Osmolar Concentration; D010447:Peptide Hydrolases / Q000378:metabolism*; D011480:Protease Inhibitors; D011817:Rabbits; D051381:Rats; D013045:Species Specificity; D013737:Testis / Q000378:metabolism; D013950:Thymus Gland / Q000378:metabolism
D002843:Chromatin; D006657:Histones; D011480:Protease Inhibitors; D010447:Peptide Hydrolases