Effects of thrombin on washed, human platelets: changes in the subcellular fractions.
Effects of thrombin on washed, human platelets: changes in the subcellular fractions.
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凝血酶对洗涤的人血小板的影响：亚细胞分数的变化。
DOI：
10.1016/0304-4165(75)90006-9
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发表时间：
1975
期刊：
影响因子：
--
通讯作者：
I. Hagen
中科院分区：
文献类型：
--
作者：
I. Hagen
已对清洗后的人血小板和用凝血酶处理的血小板进行压力均质化和亚细胞分级分离，以进行所谓的释放反应。电子显微镜显示，从对照样品中获得的颗粒区域对应于膜囊泡（B）、小储存颗粒（D）以及线粒体和较大储存颗粒（E）。在从凝血酶处理的血小板中分离的颗粒区中只能观察到少量的储存颗粒。蔗糖梯度图谱的目视比较显示，凝血酶处理的样品中有一个颗粒组分（D）消失了。wt 145 500 ± 1000）在凝血酶处理的样品的细胞外相（除去血小板后的上清液）和对照的颗粒级分（D和E）中（mol. wt 147000 ± 1000）。用凝血酶在37°C孵育洗涤过的完整血小板5分钟，然后对分离的膜部分进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，结果显示，与从对照血小板分离的膜相比，蛋白质带型没有明显差异。然而，凝血酶处理30分钟后，一条蛋白条带（mol. wt 183000 ± 3500消失。测定了蛋白质和β-N-乙酰氨基葡萄糖苷酶活性在亚细胞组分中的分布。两者主要在可溶性级分中回收（>77%）。对照品的颗粒级分D和E含有梯度中β-N-乙酰氨基葡糖苷酶总量的30% ± 0.8%和6.4% ± 1.3%。桑色素处理的细胞的组分E含有总量的3.3% ± 1.0%，而组分D缺乏。
Pressure homogenization and subcellular fractionation has been performed on washed, human platelets and platelets treated with thrombin to undergo the so-called release reaction. Electron microscopy revealed that the particulate zones obtained from the control sample corresponded to membrane visicles (B), small storage granules (D) as well as mitochondria and larger storage granules (E). Only a few storage granules could be observed in the particulate zones isolated from thrombin-treated platelets. Visual comparison of the sucrose gradient patterns revealed that one granule fraction (D) had dissappeared from the thrombin-treated sampl.Sodium dodecyclysulfate-polyacrylamide gel electrophoresis showed a major protein band (mol. wt 145 500 ± 1000) in the extracellular phase (supernatant after removal of the platelets) of the thrombin-treated sample and in the granule fractions (D and E) of the control (mol. wt 147 000 ± 1000). Incubation of whole, washed platelets with thrombin for 5 min 37°C followed by sodium dodecylsulfate-polyacrylamide gel electrophoresis of the isolated membrane fraction revealed no reporducible differences in the protein band pattern compared to menbrane isolated from control platelets. However, after treatment with thrombin for 30 min, a protein band (mol. wt 183 000 ± 3500 had disappeared.The distribution of protein andβ-N-acetylgluciosaminidase activity among the subcellular fractions were measured. Both were mainly recovered in the soluble fraction (>77%). The granule fractions, D and E of the control contained 30% ± 0.8% and 6.4% ± 1.3% of the total amount ofβ-N-acetylglucosaminidase in the gradient. Fraction E of the thormbin-treated cells contained 3.3% ± 1.0% of total while fraction D was lacking.