1887

Abstract

Surmmary

Cell-free incorporation of amino acid into paramecium protein was accomplished by using ribosomes, soluble fraction, guanosine triphosphate and adenosine triphosphate. Less than 20% of the incorporated label was detected in the soluble fraction, indicating that little if any complete synthesis occurred. Incorporation was markedly decreased by submicrogram concentrations of RNase and by decreasing [Mg] to below 3 μmoles/ ml. A ‘pH 5 fraction’ from mouse liver was able to replace the paramecium soluble fraction but attempts to obtain active ‘pH 5 fractions’ from paramecium failed. Evidence is presented for the presence of polyribosomes in paramecium; there was some indication that they were active in amino acid incorporation. Following incorporation, 80s ribosomes labelled with amino acid were recovered; subjecting these to Mg-deficient buffer caused dissociation into 45s and 30s units. A considerable portion of the label remained with the heavier unit.

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-47-1-1
1967-04-01
2021-08-04
Loading full text...

Full text loading...

/deliver/fulltext/micro/47/1/mic-47-1-1.html?itemId=/content/journal/micro/10.1099/00221287-47-1-1&mimeType=html&fmt=ahah

References

  1. Allen E. H., Schweet R. S. 1962; Synthesis of hemoglobin in a cell-free system I. Properties of the complete System. J. biol. Chem. 237:760
    [Google Scholar]
  2. Allen D. W., Zamecnik P. C. 1962; The effect of puromycin on rabbit reticulocyte ribosomes. Biochim.biophys. Acta 55:865
    [Google Scholar]
  3. Campbell P. N. 1965; The biosynthesis of proteins. Progress in Biophysics and Molecular Biology 15:3
    [Google Scholar]
  4. Chesters J. K. 1966; Protein synthesis by cell-free extracts of Crithidiaoncopelti.. Biochim.biophys. Acta 114:385
    [Google Scholar]
  5. Fruton J. S. 1963; Chemical aspects of protein synthesis.. In The Proteins, second edition. ed. by Neurath H. vol. i: p. 189 New York: Academic Press;
    [Google Scholar]
  6. Lowry O. H., Rosebrough N. J., Farr A. L., Randall R. J. 1951; Protein measurement with the folin phenol reagent. J. biol. Chem. 193:265
    [Google Scholar]
  7. Macindoe H., Reisner A. H. 1967; Adsorption titration as a specific semi-quantitative assay for soluble and bound paramecium serotypic antigen. Aust. J. Biol. Sci. (in the Press).
    [Google Scholar]
  8. Mager J. 1960; Chloramphenicol and chlortetracycline in inhibition of amino acid incorporation into proteins in a cell-free system from Tetrahymenapyriformis.. Biochim.biophys. Acta 38:150
    [Google Scholar]
  9. Mager J., Lipmann F. 1958; Amino acid incorporation and the reversion of its initial phase with cell-free Tetrahymena preparations. Proc. natn.Acad. Sci. U.S.A. 44:305
    [Google Scholar]
  10. Marcus A., Feeley J. 1965; Protein synthesis in imbibed seeds. II. Polysome formation during imbibition. J. biol. Chem. 240:1675
    [Google Scholar]
  11. Matthaei J. H., Nirenberg M. W. 1961; Characters and stabilization of DNAse-sensitive protein synthesis in Escherichia coli extracts. Proc. natn.Acad. Sci. U.S.A. 47:1580
    [Google Scholar]
  12. Nomoto M., Narahashi Y., Murakami M. 1960; The proteolytic enzyme of Streptomyces griseus. Part 7: Substrate specificity of S. griseus and protease. J. Biochem. 48:906
    [Google Scholar]
  13. Orange M., Rhein H. C. 1951; Microestimation of magnesium in body fluids. biol. Chem. 189:379
    [Google Scholar]
  14. Philipps G. R. 1966; Protein synthesis on reticulocyte ribosomes. II. The binding of peptidyl-sRNA to ribosomes. J. mol. Biol. 15:587
    [Google Scholar]
  15. Rendi R., Ochoa S. 1962; Effect of chloramphenicol on protein synthesis in cell-free preparations of Escherichia coli.. J. biol. Chem. 237:3711
    [Google Scholar]
  16. Seaman G. R. 1962; Protein synthesis by kinetosomes isolated from the protozoan Tetrahymena. Biochim.biophys. Acta 55:889
    [Google Scholar]
  17. So A. G., Davie W. E. 1963; The incorporation of amino acids into protein in a cell-free system from yeast. Biochemistry 2:132
    [Google Scholar]
  18. Tashiro Y., Siekevitz P. 1965; Localization on hepatic ribosomes of protein newly synthesized in vivo. J. mol. biol. 11:166
    [Google Scholar]
  19. Zamecnik P. C. 1962; Unsettled question in the field of protein synthesis. Biochem. J. 85:257
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-47-1-1
Loading
/content/journal/micro/10.1099/00221287-47-1-1
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error