POLYRIBOSOME SIZE ANALYSIS - MEASUREMENT OF NUMBER AVERAGE POLYRIBOSOME SIZES

被引：11

作者：

FIORETTI, WC

DAVIS, DF

LEDFORD, BE

机构：

[1] Department of Biochemistry, Medical University of South Carolina, Charleston, SC 29403

来源：

BIOCHIMICA ET BIOPHYSICA ACTA | 1979年 / 564卷 / 01期

关键词：

Albumin synthesis; Isokinetic gradient centrifugation; Polyribosome size; Translational analysis;

D O I：

10.1016/0005-2787(79)90190-4

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

The analysis of translational efficiencies of specific mRNAs requires a determination of the polyribosome size. The appropriate value to use in such calculations is the number-average size. A method is described for accurately measuring the number-average size of total and of specific protein synthesizing polyribosomes using isokinetic sucrose density gradients and 125I-labeled antibodies. By this method, we demonstrated that albumin synthesizing polyribosomes from a serum albumin secreting mouse hepatoma cell line exist over a broad range from trimers to 20-mers (mean 6-10). The specificity of antibody interaction with polyribosomes was demonstrated using cells not synthesizing mouse serum albumin, and by demonstrating that 125I-anti ovalbumin does not bind to mouse hepatoma polyribosomes. Treatment of the mouse hepatoma cells with 1 μM cycloheximide shifted practically all of the monomers into polyribosomes resulting in an increase in the number-average size of the albumin synthesizing polyribosomes. Cycloheximide treatment, however, did not eliminate the size heterogeneity in the albumin synthesizing polyribosomes. © 1979.

引用

页码：79 / 89

页数：11

共 19 条

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