Occurrence and particle character of aminoacyl-tRNA synthetases in the post-microsomal fraction from rat liver.
Occurrence and particle character of aminoacyl-tRNA synthetases in the post-microsomal fraction from rat liver.
复制标题
大鼠肝脏微粒体后部分中氨酰基-tRNA 合成酶的出现和颗粒特征。
DOI：
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发表时间：
1972
期刊：
影响因子：
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通讯作者：
H. Bloemendal
中科院分区：
文献类型：
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作者：
C. Vennegoor;H. Bloemendal
以105000 ×g离心15 h，从大鼠肝细胞液中分离出一种微粒体后组分（组分X）。许多氨酰-tRNA合成酶的活性在所得沉淀中比在上清液中高得多。这些酶（谷氨酰胺基-、异亮氨酰-、亮氨酰-、赖氨酰-和甲硫氨酰-tRNA合成酶）通过Sephadex G-200凝胶过滤、DEAE-Sephadex A-50和羟基磷灰石层析进行纯化。与粗细胞液相比，纯化倍数达到120至170倍。
在所有纯化步骤中，包括在蔗糖梯度上的额外离心或等电聚焦，五种酶的活性的主峰重合。
在电子显微镜中，可以看到尺寸为11-14 nm的圆形和矩形颗粒。从这些研究结果中，我们认为，在大鼠肝细胞，如果不是在所有的动物细胞，部分氨酰-tRNA合成酶发生在颗粒状态。
A post-microsomal fraction (Fraction X) was isolated from cell sap from rat liver by centri-fugation at 105000 ×g for 15 h. The activity of a number of aminoacyl-tRNA synthetases was considerably higher in the resulting pellet than in the supernatant. These enzymes (glutaminyl-, isoleucyl-, leucyl-, lysyl-, and methionyl-tRNA synthetase) were purified by gel filtration on Sephadex G-200, chromatography on DEAE-Sephadex A-50 and on hydroxyapatite. As compared with crude cell sap a 120- to 170-fold purification was achieved.
In all purification steps, including additional centrifugation on sucrose gradients or isoelectric focusing, the major peak of activity of the five enzymes coincided.
In the electron microscope circular and rectangular particles, 11–14 nm in size, could be visualized. From these findings we suggest that in rat liver cells, if not in all animal cells, part of the aminoacyl-tRNA synthetases occur in a particulate state.