A comparison of methods for the isolation and fractionation of reticulocyte ribosomes

1. Polysomes, ribosomes and pH5 enzymes were isolated from rabbit reticulocytes by acidifying the post-mitochondrial supernatant to pH6.0 to precipitate all ribonucleoprotein particles and about half the pH5 enzymes; the precipitate was redissolved in buffer, pH7.6, and fractionated by zone centrifu...

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Bibliographic Details
Published in:Biochemical journal 1965-08, Vol.96 (2), p.500-506
Main Authors: Arnstein, H R, Cox, R A, Gould, H, Potter, H
Format: Article
Language:English
Subjects:
Animals
Chemical Phenomena
Chemistry
Chemistry Techniques, Analytical
Edetic Acid
Hydrogen-Ion Concentration
In Vitro Techniques
Magnesium
Rabbits
Reticulocytes
Ribosomes
RNA, Messenger
Ultracentrifugation
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Summary:1. Polysomes, ribosomes and pH5 enzymes were isolated from rabbit reticulocytes by acidifying the post-mitochondrial supernatant to pH6.0 to precipitate all ribonucleoprotein particles and about half the pH5 enzymes; the precipitate was redissolved in buffer, pH7.6, and fractionated by zone centrifuging. 2. The isolation of polysome-rich and ribosome-rich fractions from the post-mitochondrial supernatant was also examined. 3. Studies of the stability of polysomes revealed that dissociation into sub-units occurred when both bound and free Mg(2+) was chelated by EDTA or when the pH was increased above pH8.8.
ISSN:0264-6021
1470-8728
DOI:10.1042/bj0960500