豚鼠乳腺泌乳期蛋白质生物合成的研究。切片和无细胞体系中α-乳白蛋白及总蛋白合成的特性。
Studies on the biosynthesis of protein by lactating guinea-pig mammary gland. Characteristics of the synthesis of alpha-lactalbumin and total protein by slices and cell-free systems.
作者信息
Brew K, Campbell P N
出版信息
Biochem J. 1967 Jan;102(1):265-74. doi: 10.1042/bj1020265.
Abstract
- Slices of lactating guinea-pig mammary gland were incubated with radioactive amino acids and the various subcellular fractions separated by centrifugation after disruption of the cells by mincing and homogenization. The most active fraction for protein synthesis appeared to be the ;mitochondrial'. 2. When the subcellular fractions were prepared without previous incubation of the cells and were then incubated with radioactive amino acid and an energy-generating system, the ;mitochondrial fraction' was at least as active for protein synthesis as the ;microsomal fraction'. 3. The ribosomes in the microsomal fraction are mainly unattached to membrane whereas those in the mitochondrial fraction are probably attached to fragments of the rough-surfaced endoplasmic reticulum. This latter fraction contains few mitochondria. 4. The combined mitochondrial and microsomal fractions incorporated radioactive amino acids into alpha-lactalbumin. 5. The radioactive leucine isolated from tryptic and chymotryptic peptides of alpha-lactalbumin synthesized in the cell-free system was not of uniform specific radioactivity. This was consistent with the polypeptide being assembled by the sequential addition of amino acids. 6. Evidence is presented for the polypeptide chain of alpha-lactalbumin being assembled from the N-terminus and for chain initiation in the cell-free system. 7. It is concluded that cell-free extracts of lactating mammary gland synthesize alpha-lactalbumin.
摘要
- 将泌乳豚鼠乳腺切片与放射性氨基酸一起孵育，通过切碎和匀浆使细胞破碎后，经离心分离出各种亚细胞组分。蛋白质合成最活跃的组分似乎是“线粒体”组分。2. 当制备亚细胞组分时不预先孵育细胞，然后将其与放射性氨基酸和能量产生系统一起孵育，“线粒体组分”在蛋白质合成方面至少与“微粒体组分”一样活跃。3. 微粒体组分中的核糖体主要未附着于膜，而线粒体组分中的核糖体可能附着于糙面内质网片段。后一组分含有的线粒体很少。4. 线粒体和微粒体组分的混合物将放射性氨基酸掺入α-乳白蛋白中。5. 从无细胞系统中合成的α-乳白蛋白的胰蛋白酶和糜蛋白酶肽段中分离出的放射性亮氨酸，其比放射性并不均匀。这与多肽通过氨基酸的顺序添加而组装一致。6. 有证据表明α-乳白蛋白的多肽链从N端开始组装，且在无细胞系统中有链起始现象。7. 得出的结论是，泌乳乳腺的无细胞提取物合成α-乳白蛋白。
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