HESS E L, LAGG S E
J Biophys Biochem Cytol. 1958 Nov 25;4(6):717-25. doi: 10.1083/jcb.4.6.717.
The extraction behavior of thymus and the composition of fractions prepared from this organ has been studied. Sequential extraction methods using 0.15 M NaCl followed by water gave information with respect to the weight fraction of cytoplasmic and nuclear constituents. Lipide, nucleic acid, and electrophoretic analysis of the extracts provided additional information. A less complex electrophoretic pattern was obtained from subsequent extracts in the sequence. Sucrose and saline dispersates obtained from tissue fragmented with either the Potter-Elvehjem homogenizer or in a Waring blendor were fractionated, using standard differential sedimentation methods. The fractions obtained by means of four different dispersion procedures were compared in terms of yield, chemical analysis, and electrophoretic composition. The quantity of material in thymus having the sedimentation characteristics of liver mitochondrial and microsomal fractions was remarkably small. Both the suspension medium employed and the method used to bring about a disruption of the cells in the tissue affected the yield of "particulate" material. The components present in the later extracts in the sequence, E(4) to E(7), in the case of sequential extraction study resembled with respect to chemical composition and electrophoretic characteristics, the microsome fraction prepared by differential sedimentation methods. About 76 per cent of the PNA in the tissue appeared to be in the cytoplasm. The remaining 24 per cent PNA was found in the nucleus and accounted for 1.7 per cent of nucleus on a dry weight basis. From 75 to 88 per cent of cytoplasmic PNA was extracted from the tissue and 76 to 94 per cent of the PNA in the extract was found in the final supernatant solutions, depending upon the dispersion methods and suspension medium used in the extraction procedure. The composition of the final supernatant fractions using differential sedimentation methods were comparable in terms of electrophoretic properties, protein concentration, nucleic acid content, and fractionation behavior to saline extracts E(1) to E(3), of thymus used in earlier studies.
对胸腺的提取行为以及从该器官制备的各组分的组成进行了研究。采用0.15M氯化钠随后用水的顺序提取方法，得到了关于细胞质和核成分重量分数的信息。提取物的脂质、核酸和电泳分析提供了更多信息。在该序列中后续提取物获得的电泳图谱较不复杂。使用标准差速沉降方法对用波特-埃尔维希匀浆器或韦林搅拌器破碎组织得到的蔗糖和盐分散液进行分级分离。比较了通过四种不同分散程序获得的级分的产率、化学分析和电泳组成。胸腺中具有肝线粒体和微粒体级分沉降特性的物质数量非常少。所使用的悬浮介质和使组织中细胞破碎的方法都会影响“颗粒状”物质的产率。在顺序提取研究中，序列中E(4)至E(7)的后续提取物中存在的成分，在化学组成和电泳特性方面类似于通过差速沉降方法制备的微粒体级分。组织中约76%的PNA似乎存在于细胞质中。其余24%的PNA存在于细胞核中，以干重计占细胞核的1.7%。根据提取过程中使用的分散方法和悬浮介质，75%至88%的细胞质PNA从组织中被提取出来，提取物中76%至94%的PNA存在于最终的上清液中。使用差速沉降方法得到的最终上清级分的组成，在电泳性质、蛋白质浓度、核酸含量和分级行为方面，与早期研究中使用的胸腺盐提取物E(1)至E(3)相当。
