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MPC - 11细胞中核糖体与重度粗面和轻度粗面内质网的膜结合情况的差异。

Differences in the association of ribosomes with heavy rough and light rough endoplasmic reticulum membranes of MPC-11 cells.

作者信息

Pryme I F

机构信息

Department of Biochemistry, University of Bergen, Norway.

出版信息

Biochim Biophys Acta. 1988 Nov 17;967(2):218-23. doi: 10.1016/0304-4165(88)90012-8.

Abstract

The treatment of total endoplasmic reticulum membranes of mouse plasmacytoma cells with EDTA resulted in an abolition of the heavy rough (HR) subfraction, while there was a large increase in smooth (S) membranes. When HR and light rough (LR) endoplasmic reticulum membranes were treated individually with EDTA and re-centrifuged on discontinuous sucrose gradients it was observed that HR were converted into S membranes, i.e. membranes virtually devoid of ribosomes. LR membranes were not affected to the same extent but there was a shift to a somewhat lower density. A quantitation of ribosomes released by EDTA showed that 95% of 60 S and 72% of 40 S subunits were removed from HR membranes while for LR membranes the corresponding values were 8.5 and 22.6% respectively. Ratios of radioactivity to absorbance at 260 nm calculated for 40 S and 60 S subunits isolated from HR and LR membranes show that 60 S subunits from LR membranes, in contrast to those from HR membranes, equilibrate only slowly with the free pool of ribosomal subunits. The results indicate that the ribosomes associated with HR membranes are 'loosely bound' and those with LR membranes 'tightly bound'. When poly(A)-containing mRNA isolated from HR and LR membranes was translated in vitro and the products analysed for light-chain immunoglobulin content, it was found that the HR fraction was enriched in light-chain mRNA.

摘要

用乙二胺四乙酸(EDTA)处理小鼠浆细胞瘤细胞的总内质网膜,导致重糙面(HR)亚组分消失,而光面(S)膜大幅增加。当分别用EDTA处理HR和轻糙面(LR)内质网膜,并在不连续蔗糖梯度上重新离心时,观察到HR膜转变为S膜,即几乎不含核糖体的膜。LR膜受影响程度较小,但密度略有下降。对EDTA释放的核糖体进行定量分析表明,95%的60S和72%的40S亚基从HR膜中被去除,而对于LR膜,相应的值分别为8.5%和22.6%。从HR和LR膜中分离出的40S和60S亚基在260nm处的放射性与吸光度之比表明,与HR膜中的60S亚基不同,LR膜中的60S亚基与核糖体亚基的游离池达到平衡的速度较慢。结果表明,与HR膜相关的核糖体是“松散结合”的,而与LR膜相关的核糖体是“紧密结合”的。当从HR和LR膜中分离出的含多聚腺苷酸(poly(A))的信使核糖核酸(mRNA)在体外进行翻译,并对产物进行轻链免疫球蛋白含量分析时,发现HR组分富含轻链mRNA。

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