SIEKEVITZ P, PALADE G E
J Biophys Biochem Cytol. 1958 Sep 25;4(5):557-66. doi: 10.1083/jcb.4.5.557.
DL-leucine-1-C(14) was administered by intracardiac injection to guinea pigs and its in vivo incorporation into the proteins of various pancreatic cell fractions followed over a period of 2 hours. The pancreas was homogenized in 0.88 M sucrose and fractionated by differential centrifugation to give nuclear, zymogen, mitochondrial, microsomal, postmicrosomal, and final supernatant fractions. The proteins of these fractions, obtained by precipitation with trichloroacetic acid followed by washing, were counted. The proteins of the microsomal fraction showed the highest early specific activity and were followed by those of the zymogen and mitochondrial fractions. The microsomal fraction was broken up into two subfractions: one consisting of detached RNP particles, the other representing mainly the microsomal content and membranes. The incorporation of labelled leucine into the proteins of microsomal subtractions and in those of postmicrosomal fractions was studied comparatively in the pancreas of fasted and fed guinea pigs as well as in the liver and pancreas of fasted animals. A tentative cytological picture of protein synthesis and transport based on these findings is presented.
将DL-亮氨酸-1-C(14)经心内注射给予豚鼠,并在2小时内追踪其在体内掺入各种胰腺细胞组分蛋白质中的情况。将胰腺在0.88M蔗糖中匀浆,通过差速离心分级分离,得到细胞核、酶原、线粒体、微粒体、微粒体后和最终上清液组分。通过用三氯乙酸沉淀然后洗涤获得这些组分的蛋白质,并进行计数。微粒体组分的蛋白质显示出最高的早期比活性,其次是酶原和线粒体组分的蛋白质。微粒体组分被分解为两个亚组分:一个由游离的核糖核蛋白颗粒组成,另一个主要代表微粒体内容物和膜。比较研究了禁食和喂食豚鼠胰腺以及禁食动物肝脏和胰腺中标记亮氨酸掺入微粒体亚组分蛋白质和微粒体后组分蛋白质中的情况。基于这些发现,给出了蛋白质合成和转运的初步细胞学图像。