Open AccessThe Integrity of Nuclear Proteins Following Incubation of Isolated Nuclei in vitro
Author(s) -
SELLWOOD Susan M.,
RICHES Pamela G.,
HARRAP Kenneth R.,
RICKWOOD David,
MacGILLIVRAY Alexander J.,
CAPPS Marilyn
Publication year - 1975
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1975.tb04027.x
Subject(s) - incubation , isoelectric point , tris , gel electrophoresis , microbiology and biotechnology , polyacrylamide gel electrophoresis , biochemistry , isoelectric focusing , biology , nuclear matrix , nuclear protein , electrophoresis , chromatin , chemistry , chromatography , dna , enzyme , gene , transcription factor
1 Nuclear proteins have been prepared from isolated nuclei of various rat and mouse tissues. The effect on these proteins of incubation of nuclei at 37° C, storage at –40° C and Triton washing is described. 2 Examination by one‐dimensional dodecylsulphate‐polyacrylamide gel electrophoresis indicated that incubation at 37° C brought about degradation of the Tris‐saline‐soluble proteins of nuclei prepared from rat liver, spleen and kidney. 3 Two‐dimensional electrophoresis, combining isoelectric focusing and dodecylsulphate electrophoresis, indicated a similar effect on the Tris‐saline‐soluble nuclear proteins of mouse liver; degradation affected all protein species. 4 Examination of histones and non‐histone chromatin proteins from mouse liver by two‐dimensional electrophoresis showed these protein groups to be stable during incubation of isolated nuclei at 37°C. 5 Storage of rat liver nuclei at –40° C and omission of routine Triton washing during preparation of nuclei were found to have little effect on the Tris‐saline‐soluble proteins, all degradation being a consequence of the subsequent incubation at 37°C. 6 Degradation of this same group of proteins did not occur during incubation of isolated nuclei from a rat ascites tumour or regenerating rat liver, 16 h after partial hepatectomy.