The isolation of Saccharomyces cerevisiae nuclear membranes with nuclease and high-salt treatment |
| |
Authors: | Karlheinz Mann Dieter Mecke |
| |
Affiliation: | Physiologisch-chemisches Institut der Universität, Hoppe-Seyler-Strasse 1, D-7400 Tübingen F.R.G. |
| |
Abstract: | Saccharomyces cerevisiae nuclear membranes were prepared from isolated nuclei by digesting chromatin with deoxyribonuclease and ribonuclease, washing of residual nuclei with 0.5 M MgCl2, and discontinuous gradient centrifugation in buffered Ficoll solutions. Electron microscopic examination of the preparations showed single membrane and double membrane vesicles and membrane sheets. Pores or residual pores were often visible. In double membrane profiles the two unit membranes were often separated by the remains of the perinuclear cistern. The nuclear membrane fragments contained 58% protein, 23.8% phospholipid, 6% sterols, 7.1% neutral acylglycerols, 4.8% RNA, and 0.3% DNA. The phospholipid content of the membrane preparations was influenced by a phospholipase activity with acidic pH optimum. |
| |
Keywords: | Nuclear membrane isolation Nuclease digestion High-salt treatment Ficoll gradient centrifugation Electron microscopy (S. cerevisiae) |
本文献已被 ScienceDirect 等数据库收录! |
|