Albi E, Viola-Magni M P
Institute of General Pathology, University of Perugia, Italy.
Cell Biol Int. 1997 Apr;21(4):217-21. doi: 10.1006/cbir.1997.0133.
Previous investigations have demonstrated the presence of phospholipids as a component of chromatin; however the mechanism of their synthesis, namely if they are synthesized in the nuclei or in the cytoplasm (microsomal fraction), from where they may eventually be transported to the nucleus, has not yet been clarified. The phosphatidylcholine, for example, can be formed, albeit in a limited amount, by an interconversion reaction between bases. The aim of the present research was to ascertain the presence of the enzyme complex responsible for this reaction in hepatocyte nuclei and in isolated nuclear membrane. The incorporation of [14C]-choline in phosphatidylcholine was assayed in microsomes, hepatocyte nuclei, liver nuclei and nuclear membranes of rat liver. The reaction was Ca(2+)-dependent and the specific activity was higher in microsomes but was present, albeit at a low level, also in nuclei and in nuclear membranes. Possible contaminations were excluded by specific microsomal markers and by the reaction time course. In fact, the nuclear reaction reached the maximum level slowly with respect to microsomes. Since the phosphatidylcholine extracted from the nuclei show an enrichment in unsaturated fatty acids of monoenoic fraction, such as oleic acid, the difference in reaction kinetics has been tentatively explained as due to the phosphatidylcholine fatty acid content. The presence of this base exchange enzyme complex may allow a fast change in chromatin phospholipid composition.
先前的研究已证明磷脂作为染色质的一种成分存在;然而,其合成机制,即它们是在细胞核中还是在细胞质(微粒体部分)中合成,最终又从何处转运至细胞核,尚未阐明。例如,磷脂酰胆碱可通过碱基间的相互转化反应形成,尽管量有限。本研究的目的是确定肝细胞核和分离的核膜中负责该反应的酶复合物的存在情况。在大鼠肝脏的微粒体、肝细胞核、肝细胞核和核膜中检测了[14C] - 胆碱掺入磷脂酰胆碱的情况。该反应依赖于Ca(2+),微粒体中的比活性较高,但在细胞核和核膜中也有存在,尽管水平较低。通过特定的微粒体标记物和反应时间进程排除了可能的污染。事实上,与微粒体相比,核反应达到最大水平的速度较慢。由于从细胞核中提取的磷脂酰胆碱在单烯酸部分的不饱和脂肪酸(如油酸)中富集,反应动力学的差异暂被解释为是由于磷脂酰胆碱的脂肪酸含量所致。这种碱基交换酶复合物的存在可能使染色质磷脂组成快速变化。