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PMID: 4953780 Published · ppublish English Journal Article

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

The Biochemical journal ·Vol. 96 ·No. 2 ·1965-08-00 ·Pages 500-6

Arnstein HR, Cox RA, Gould H, Potter H

Abstract

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.

MeSH Terms
Animals Chemical Phenomena Chemistry Chemistry Techniques, Analytical Edetic Acid Hydrogen-Ion Concentration In Vitro Techniques Magnesium RNA, Messenger Rabbits Reticulocytes Ribosomes Ultracentrifugation
Chemicals
RNA, Messenger Edetic Acid Magnesium
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Arnstein H R
Cox R A
Gould H
Potter H
References (10)
10 references, click to expand
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  2. Ribosomal aggregate engaged in protein synthesis: characterization of the ergosome.
    Nature. 1963 Feb 2;197:430-5 PMID: 14000153
  3. Unstable ribonucleic acid revealed by pulse labelling of Escherichia coli.
    Nature. 1961 May 13;190:581-5 PMID: 13708983
  4. Electron Microscope Studies of Ribosomal Clusters Synthesizing Hemoglobin.
    Science. 1962 Dec 28;138(3548):1399-403 PMID: 17753859
  5. Studies on ribosomes from reticulocytes.
    Biochim Biophys Acta. 1961 Apr 29;49:113-29 PMID: 13778314
  6. Ultracentrifugal studies on ribonucleoprotein from rat liver microsomes.
    J Biol Chem. 1959 Jun;234(6):1441-6 PMID: 13654394
  7. A multiple ribosomal structure in protein synthesis.
    Proc Natl Acad Sci U S A. 1963 Jan 15;49:122-9 PMID: 13998950
  8. Factors affecting protein synthesis in vitro in rabbit reticulocytes.
    J Biol Chem. 1957 Dec;229(2):1059-70 PMID: 13502365
  9. Function of polyuridylic acid and ribonucleic acid in protein biosynthesis by ribosomes from mammalian reticulocytes.
    Nature. 1962 Jun 16;194:1042-4 PMID: 13862467
  10. THE SYNTHESIS OF HEMOGLOBIN IN A CELL-FREE SYSTEM.
    Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1029-35 PMID: 16590302
Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
Published
1965-08-00
Pages
500-6
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1207066
Subset
IM
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