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PMID: 4429540 Published · ppublish English Journal Article

Changes in free and membrane-bound ribosomes during the development of chick liver. A new cell-fractionation approach.

The Biochemical journal ·Vol. 138 ·No. 3 ·1974-03-00 ·Pages 359-71

O'Toole K, Pollak JK

Abstract

A major difficulty in studying quantitative changes in free and membrane-bound ribosomes in a tissue under different physiological conditions is that membrane-bound ribosomes are not usually recovered quantitatively in a conventional microsomal fraction. This problem was resolved for developing chick liver by determining the conditions for the isolation of a microsomal fraction containing the highest practicable yield of rough vesicles, and then separating it into free-ribosome- and rough-vesicle-containing fractions. With the aid of a marker enzyme for the microsomal membranes and the RNA content of the recovered membrane-bound ribosomes, it was possible to correct for the recovery of rough vesicles and hence to determine the concentration of membrane-bound ribosomes in the homogenate. Despite the fact that morphological studies have suggested that most of the cellular ribosomes are not bound to membrane in chick liver cells at the earliest developmental age studied (6 days of egg incubation), 49% of the total ribosomes were found to be membrane-bound by using the new fractionation technique. This fraction increased (to 66%) during development. The discrepancy between the cell-fractionation and morphological approaches could not be attributed to artifacts of the separation method but rather to difficulties inherent in the morphological approach.

MeSH Terms
Age Factors Animals Cell Fractionation Centrifugation, Density Gradient Chick Embryo Chickens Liver/embryology,growth & development Membranes/metabolism Microscopy, Electron Mitochondria, Liver/analysis,ultrastructure RNA, Ribosomal/analysis Ribosomes/analysis,metabolism Ultracentrifugation
Chemicals
RNA, Ribosomal
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
O'Toole K
Pollak J K
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55 references, click to expand
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj1380359
Published
1974-03-00
Pages
359-71
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1166221
Subset
IM
External Links
PubMed source DOI: 10.1042/bj1380359 PMC Full Text View journal details
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