Chromatin-nuclear envelope complex from rat liver: isolation and purification |
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| Authors: | A N Prusov D Fais V Y Polyakov B F Vanyushin |
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| Affiliation: | 1. Anti-Cancer Drug Section, Department of Drug Research and Medicine Evaluation, Istituto Superiore di Sanità (National Institute of Health), Rome, Italy;2. Institut de Recherche Expérimentale et Clinique (IREC), Pole of Pharmacology, Université Catholique de Louvain (UCL), Brussels, Belgium;3. Department of Gastroenterology, Hannover Medical School, Hannover, Germany;4. Oral Medicine, Oral Surgery and Implantology Unit, Faculty of Medicine and Dentistry, Instituto de Investigación Sanitaria de Santiago (IDIS), Santiago de Compostela, Spain;5. Cancer Center Karolinska, Department of Oncology-Pathology, Karolinska Institute, Stockholm, Sweden;6. Laboratory for Orthopaedic Pathophysiology and Regenerative Medicine, Istituto Ortopedico Rizzoli, Bologna, Italy;7. Instituto de Biología Clínica y Metabolismo (IBCM), (Postas) Vitoria, Spain |
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| Abstract: | We describe a method for the isolation of a fraction of nuclear envelope (NE) from rat liver. The method includes mild treatment of pure nuclei with either endonuclease of DNase I under low ionic strength conditions in the presence of magnesium, which allows the nucleomeric organization of the chromatin (Ch) to be preserved. The NEs were purified by centrifugation in sucrose gradients followed by floatation in sucrose. No more than 3% of the Ch present in the purified Ch-NE complexes was due to the non specific adsorption of Ch to the NE. The main components of the complex (Ch and NE) retained their in situ ultrastructure. The complex consisted of 9--10% DNA, 3--4% RNA, about 63% protein and about 24% phospholipids. |
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