Nuclear membranes from mammalian liver. I. Isolation procedure and general characterization.
Nuclear membranes were isolated from rat and pig liver by sonication of highly purified nuclear fractions and subsequent removal of adhering nucleoproteins in a high salt medium. The fractions were examined in the electron microscope by both negative staining and thin sectioning techniques and were found to consist of nuclear envelope fragments of widely varying sizes. Nuclear pore complex constituents still could frequently be recognized. The chemical composition of the nuclear membrane fractions was determined and compared with those of microsomal fractions prepared in parallel. For total nuclei as well as for nuclear membranes and microsomes, various enzyme activities were studied. The results indicate that a similarity exists between both fractions of cytomembranes, nuclear envelope, and endoplasmic reticulum, with respect to their RNA:protein ratio and their content of polar and nonpolar lipids. Both membranous fractions had many proteins in common including some membrane-bound enzymes. Activities in Mg-ATPase and the two examined cytochrome reductases were of the same order of magnitude. The content of cytochrome b(5) as well as of P-450 was markedly lower in the nuclear membranes. The nuclear membranes were found to have a higher buoyant density and to be richer in protein. The glucose-6-phosphatase and Na-K-ATPase activities in the nuclear membrane fraction were very low. In the gel electrophoresis, in addition to many common protein bands, some characteristic ones for either microsomal or nuclear membranous material were detected. Significant small amounts of DNA and RNA were found to remain closely associated with the nuclear envelope fragments. Our findings indicate that nuclear and endoplasmic reticulum membranes which are known to be in morphological continuity have, besides a far-reaching similarity, some characteristic differences.
D000135:Acid Phosphatase / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D002467:Cell Nucleus / Q000032:analysis* / Q000201:enzymology; D003580:Cytochromes / Q000378:metabolism; D004247:DNA / Q000032:analysis; D004589:Electrophoresis, Disc; D005260:Female; D005952:Glucose-6-Phosphatase / Q000378:metabolism; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000166:cytology*; D008297:Male; D008566:Membranes / Q000032:analysis; D008854:Microscopy, Electron; D008862:Microsomes, Liver / Q000032:analysis / Q000201:enzymology; D010743:Phospholipids / Q000032:analysis; D011506:Proteins / Q000032:analysis; D012313:RNA / Q000032:analysis; D051381:Rats; D013045:Species Specificity; D013552:Swine
D003580:Cytochromes; D010743:Phospholipids; D011506:Proteins; D012313:RNA; D004247:DNA; D005969:Glutamate Dehydrogenase; D000469:Alkaline Phosphatase; D000135:Acid Phosphatase; D005952:Glucose-6-Phosphatase; D000251:Adenosine Triphosphatases