Slaby F, Brown C
J Cell Biol. 1974 Jun;61(3):613-32. doi: 10.1083/jcb.61.3.613.
The equilibrium density distribution, protein composition, and secretory character of mouse mammary epithelial rough microsomes have been determined during differentiation. The density range exhibited by the rough microsomes broadens during mammary development; rough microsomes within the 1.25-1.29 g/ml density range appear soon after conception and then within the 1.30-1.34 range after the onset of lactation. The appearance of these denser microsomes represents the progressive increase of the average ribosome content of the rough endoplasmic reticulum (ER) during gestation and lactation. Fractionation of rough microsomal proteins by sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveals that two proteins, having molecular weights of 57,000 and 76,000, occur to a significant extent only during lactation and are then most prominent in the very dense rough microsomes of the 1.30-1.34 range. Nascent polypeptide chains discharged (by incubation with puromycin) from 17-days lactation rough microsomes in either the 1.21-1.29 or 1.30-1.34 density range are distributed equally between the intra- and extravesicular compartments. Whereas 36% of the chains are discharged intravesicularly from 1-day lactation rough microsomes in the 1.30-1.34 range, only 25% are so discharged from those in the 1.21-1.29 range. The results indicate (a) that there is no correlation between the relative levels in lactation rough microsomes of the two microsomal proteins which become prominent during lactation and the extent of secretory activity and (b) that for a short period after parturition the rough ER elements bearing high surface densities of ribosomes have a greater proportion of ribosomes synthesizing milk proteins than the rough ER elements with moderate ribosome densities.
在分化过程中,已对小鼠乳腺上皮粗面微粒体的平衡密度分布、蛋白质组成和分泌特性进行了测定。在乳腺发育过程中,粗面微粒体呈现的密度范围会变宽;在受孕后不久就会出现密度范围在1.25 - 1.29克/毫升的粗面微粒体,而在泌乳开始后则会出现密度范围在1.30 - 1.34的粗面微粒体。这些密度更高的微粒体的出现代表着在妊娠期和泌乳期粗面内质网(ER)平均核糖体含量的逐渐增加。通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳对粗面微粒体蛋白质进行分级分离显示,两种分子量分别为57,000和76,000的蛋白质仅在泌乳期大量出现,并且在密度范围为1.30 - 1.34的非常致密的粗面微粒体中最为突出。从17天泌乳期的粗面微粒体(密度范围为1.21 - 1.29或1.30 - 1.34)中通过与嘌呤霉素孵育释放出的新生多肽链,在囊泡内和囊泡外区室之间均匀分布。而在密度范围为1.30 - 1.34的1天泌乳期粗面微粒体中,36%的链从囊泡内释放,在密度范围为1.21 - 1.29的粗面微粒体中,只有25%的链是这样释放的。结果表明:(a)在泌乳期粗面微粒体中,在泌乳期变得突出的两种微粒体蛋白质的相对水平与分泌活性程度之间没有相关性;(b)在分娩后的短时间内,核糖体表面密度高的粗面内质网元件比核糖体密度适中的粗面内质网元件具有更大比例的核糖体合成乳蛋白。