Faculty Research 1980 - 1989
Specific binding of cholesterol to chromatin prepared from mouse spleen cells.
Document Type
Article
Publication Date
1984
Keywords
Cholesterol: me, Chromatin: me, Chromatography-Gel, Chromatography-Thin-Layer, Deoxyribonuclease-I: me, Lymphocytes: me, ul, Male, Mice, Mice-Inbred-C57BL, Spleen: cy, SUPPORT-U-S-GOVT-P-H-S
First Page
94
Last Page
99
JAX Source
Can-J-Biochem-Cell-Biol. 1984 Feb-Mar; 62(2-3):94-9.
Grant
CA19305
Abstract
Mouse spleen cell suspensions were incubated with tritiated cholesterol for various time intervals. The chromatin of these cells was then isolated, washed with Triton X-100, and fractionated on Sephadex columns. It was found that cholesterol binds specifically to the chromatin. The binding was saturated after 45 min of incubation and also displayed characteristics typical of dose-dependent binding. The number of cholesterol molecules bound per nucleus was estimated to be on the order of 10 000. Oxygenated sterols, such as 25-hydroxycholesterol and 7-ketocholesterol, did not compete for the binding with [3H]cholesterol if added in 20-fold molar excess to the incubation medium of the cells. Chromatographic analyses on Sephadex columns displayed a distinct peak of radioactivity. The protein-sterol complex had an apparent molecular weight of 180 000 +/- 27 000. Using extensive digestion with DNase I (EC 3.1.21.1) it could be concluded that DNA, binding to the complex, did not influence the estimate of the molecular weight, whereas digestion with pronase or treatment with sodium dodecyl sulfate destroyed the complex. Additional experiments using sucrose density gradients (5-20%) showed also, that [3H]cholesterol was bound to chromatin by one or several proteins.
Recommended Citation
Regenass KM,
Heiniger HJ.
Specific binding of cholesterol to chromatin prepared from mouse spleen cells. Can-J-Biochem-Cell-Biol. 1984 Feb-Mar; 62(2-3):94-9.