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非洲爪蟾卵巢核糖体的蛋白质

The proteins of Xenopus ovary ribosomes.

作者信息

Ford P J

出版信息

Biochem J. 1971 Dec;125(4):1091-107. doi: 10.1042/bj1251091.

DOI:10.1042/bj1251091
PMID:5004147
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1178274/
Abstract
  1. The preparation of ribosomes and ribosomal subunits from Xenopus ovary is described. 2. The yield of once-washed ribosomes (buoyant density in caesium chloride 1.601g.cm(-3); 44% RNA, 56% protein by chemical methods) was 10.1mg/g wet wt. of tissue. 3. Buoyant density in caesium chloride and RNA/protein ratios by chemical methods have been determined for ribosome subunits produced by 1.0mm-EDTA or 0.5m-potassium chloride treatment and also for EDTA subunits extracted with 0.5m-, 1.0m- or 1.5m-potassium chloride, 4. Analysis of ribosomal protein on acrylamide gels at pH4.5 in 6m-urea reveals 24 and 26 bands from small and large EDTA subunits respectively. The actual numbers of proteins are greater than this, as many bands are obviously doublets. 5. Analysis of the proteins in the potassium chloride extract and particle fractions showed that some bands are completely and some partially extracted. Taking partial extraction as an indication of possible doublet bands it was found that there were 12 and 20 such bands in the small and large subunits respectively, making totals of 36 and 46 proteins. 6. From the measured protein contents and assuming weight-average molecular weights for the proteins of large and small subunits close to those observed for eukaryote ribosomal proteins it is possible to compute the total numbers of protein molecules per particle. It appears that too few protein bands have been identified on acrylamide gels to account for all the protein in the large subunit, but probably enough for the small subunit.
摘要
  1. 本文描述了从非洲爪蟾卵巢制备核糖体及核糖体亚基的方法。2. 单次洗涤后的核糖体(在氯化铯中的浮力密度为1.601g.cm⁻³;化学方法分析RNA含量为44%,蛋白质含量为56%)产量为每克湿组织10.1mg。3. 已测定了经1.0mM - EDTA或0.5M - 氯化钾处理产生的核糖体亚基,以及用0.5M、1.0M或1.5M - 氯化钾提取的EDTA亚基在氯化铯中的浮力密度和化学方法测定的RNA/蛋白质比率。4. 在pH4.5、6M - 尿素条件下,对聚丙烯酰胺凝胶上的核糖体蛋白进行分析,结果显示小EDTA亚基和大EDTA亚基分别有24条和26条条带。由于许多条带明显是双重条带,所以实际蛋白质数量比这个多。5. 对氯化钾提取物和颗粒组分中的蛋白质进行分析表明,一些条带能被完全提取,一些则只能部分提取。以部分提取作为可能存在双重条带的指示,发现小亚基和大亚基中分别有12条和20条这样的条带,因此小亚基和大亚基的蛋白质总数分别为36种和46种。6. 根据测得的蛋白质含量,并假设大亚基和小亚基蛋白质的重均分子量接近真核生物核糖体蛋白的观测值,就可以计算出每个颗粒中蛋白质分子的总数。看起来在聚丙烯酰胺凝胶上鉴定出的蛋白质条带太少,无法解释大亚基中的所有蛋白质,但对于小亚基可能足够了。
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9978/1178274/38153ad4eb63/biochemj00640-0188-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9978/1178274/17878a34a0da/biochemj00640-0186-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9978/1178274/edc71e260d04/biochemj00640-0187-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9978/1178274/38153ad4eb63/biochemj00640-0188-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9978/1178274/17878a34a0da/biochemj00640-0186-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9978/1178274/edc71e260d04/biochemj00640-0187-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9978/1178274/38153ad4eb63/biochemj00640-0188-a.jpg

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引用本文的文献

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2
The extraction of proteins from eukaryotic ribosomes and ribosomal subunits.从真核生物核糖体及核糖体亚基中提取蛋白质。
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Characterization of chloroplast and cytoplasmic ribosomal proteins of Chlamydomonas reinhardi by two-dimensional gel electrophoresis.

本文引用的文献

1
Bacterial ribosome.细菌核糖体
Bacteriol Rev. 1970 Sep;34(3):228-77. doi: 10.1128/br.34.3.228-277.1970.
2
Protein measurement with the Folin phenol reagent.使用福林酚试剂进行蛋白质测定。
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THE CHROMOSOMAL SITE SPECIFYING A RIBOSOMAL PROTEIN IN ESCHERICHIA COLI.大肠杆菌中指定核糖体蛋白的染色体位点。
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Localization of ribosomal protein S1 in the granular component of the interphase nucleolus and its distribution during mitosis.核糖体蛋白S1在间期核仁颗粒组分中的定位及其在有丝分裂期间的分布。
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CONSERVATION OF RIBOSOMES DURING BACTERIAL GROWTH.细菌生长过程中核糖体的保守性
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RNA SYNTHESIS DURING THE DEVELOPMENT OF XENOPUS LAEVIS, THE SOUTH AFRICAN CLAWED TOAD.非洲爪蟾(非洲爪蟾)发育过程中的RNA合成。
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