Effect of divalent cations on the extraction of nuclear proteins.
TSCM-(0.01 M Tris-HCl, 0.25 sucrose, 3.8 mM CaCl2, 12 mM MgCl2, pH 7.2) and TS-(0.01 M Tris-HCl, 0.25 M sucrose, pH 7.2)-soluble proteins of isolated rat liver nuclei were analyzed by a two-dimensional polyacrylamide gel electrophoresis method. Light microscopy showed that treatment with the TSCM solution did not alter the morphology of isolated nuclei. After treatment with the divalent cation-free TS solution, the isolated nuclei were swollen; some were disrupted and clumped together. The TSCM solution extracted 20 additional proteins including histones. These results suggest that Ca++ and Mg++ play an intracellular role in the interactions between nuclear proteins and chromatin.
D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002467:Cell Nucleus / Q000187:drug effects; D002843:Chromatin / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects; D008274:Magnesium / Q000494:pharmacology*; D008297:Male; D009698:Nucleoproteins / Q000302:isolation & purification*; D051381:Rats; D012995:Solubility; D014325:Tromethamine
D002843:Chromatin; D009698:Nucleoproteins; D014325:Tromethamine; D008274:Magnesium; D002118:Calcium