Hilderson H J, De Wolf M J, Lagrou A R, Dierick W S
Biochem J. 1975 Dec;152(3):601-7. doi: 10.1042/bj1520601.
A combined mitochondrial and light mitochondrial fraction and a microsomal fraction were isolated from bovine thyroid gland and fractionated further in a B-XIV zonal rotor. A density gradient ranging from 20 to 50% (w/w) sucrose was used. The rotor was operated for 3 h at 45 000 rev./min. All manipulations were performed at 4 degrees C and at pH 7.4. 2. Membranous material was recovered in two zones: zone I, containing microsomal material derived from both smooth endoplasmic reticulum and plasma membranes and probably also from other smooth membranes; zone II, containing material from rough endoplasmic reticulum. 3. Increasing the pH of the medium up to 8.6, or the addition of Mg2+ to the medium resulted in the formation of a single zone at intermediate densities (aggregation of membranes?). An analogous effect was obtained after treatment with Pb (NO3) 2. 4. In the presence of heparin (50 i.u./ml) the bulk of the membranes was found in zone I. This was due to the release of ribosomes from the rough endoplasmic reticulum.
从牛甲状腺中分离出线粒体和轻线粒体组分以及微粒体组分,并在B-XIV区带转子中进一步分级分离。使用了20%至50%(w/w)蔗糖的密度梯度。转子在45000转/分钟下运行3小时。所有操作均在4℃和pH 7.4条件下进行。2. 膜性物质在两个区带中回收:I区,含有源自滑面内质网和质膜以及可能还来自其他滑面膜的微粒体物质;II区,含有来自粗面内质网的物质。3. 将培养基的pH提高到8.6,或向培养基中添加Mg2+,导致在中等密度处形成单个区带(膜聚集?)。用Pb(NO3)2处理后获得类似效果。4. 在肝素(50国际单位/毫升)存在下,大部分膜位于I区。这是由于核糖体从粗面内质网释放所致。