Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1972 Nov;130(1):19–25. doi: 10.1042/bj1300019

Binding of rat liver and hepatoma polyribosomes to stripped rough endoplasmic reticulum in vitro. Biological or an artifact?

A A Hochberg *, F W Stratman *, Rainer N Zahlten *, H P Morris †, H A Lardy *
PMCID: PMC1174296  PMID: 4347780

Abstract

Exposed thiol groups do not appear to be related to the binding of 32P-labelled polyribosomes to stripped rough endoplasmic reticulum in vitro. Treating stripped rough endoplasmic reticulum with GSSG did not diminish binding of polyribosomes, suggesting that binding in vitro has no correlation with the inhibition of protein synthesis in vitro reported by Kosower et al. (1971). Thiol reagents, which are known to dissociate ribosomes, did not significantly decrease binding of 32P-labelled polyribosomes to stripped rough endoplasmic reticulum. Denaturing the protein of 32P-labelled polyribosomes or stripped rough endoplasmic reticulum of liver or hepatoma with heat, trichloroacetic acid, or HClO4 did not alter the binding in vitro. Therefore, the practice of measuring the binding of 32P-labelled polyribosomes to stripped rough endoplasmic reticulum in vitro (Shires et al., 1971b) is an unsuitable indicator of biological significance in the intact cell.

Full text

PDF
19

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bermek E., Mönkemeyer H., Berg R. The role of SH-groups of human ribosomal subunits in polypeptide synthesis. Biochem Biophys Res Commun. 1971 Dec 3;45(5):1294–1299. doi: 10.1016/0006-291x(71)90159-8. [DOI] [PubMed] [Google Scholar]
  2. Blobel G., Sabatini D. Dissociation of mammalian polyribosomes into subunits by puromycin. Proc Natl Acad Sci U S A. 1971 Feb;68(2):390–394. doi: 10.1073/pnas.68.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bloemendal H., Bont W. S., de Vries M., Benedetti E. L. Isolation and properties of polyribosomes and fragments of the endoplasmic reticulum from rat liver. Biochem J. 1967 Apr;103(1):177–182. doi: 10.1042/bj1030177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blyth C. A., Freedman R. B., Rabin B. R. Sex specific binding of steroid hormones to microsomal embranes of rat liver. Nat New Biol. 1971 Mar 31;230(13):137–139. doi: 10.1038/newbio230137a0. [DOI] [PubMed] [Google Scholar]
  5. Campbell P. N. Functions of polyribosomes attached to membranes of animal cells. FEBS Lett. 1970 Mar 16;7(1):1–7. doi: 10.1016/0014-5793(70)80603-2. [DOI] [PubMed] [Google Scholar]
  6. Dice J. F., Schimke R. T. Turnover and exchange of ribosomal proteins from rat liver. J Biol Chem. 1972 Jan 10;247(1):98–111. [PubMed] [Google Scholar]
  7. Dunn A. J. The effects of sulfhydryl reagents on rat liver ribosomes and polyribosomes. Arch Biochem Biophys. 1970 Jan;136(1):203–211. doi: 10.1016/0003-9861(70)90342-5. [DOI] [PubMed] [Google Scholar]
  8. Frieden C. Protein-protein interaction and enzymatic activity. Annu Rev Biochem. 1971;40:653–696. doi: 10.1146/annurev.bi.40.070171.003253. [DOI] [PubMed] [Google Scholar]
  9. Furano A. V., Harris M. I. The activity of ribosomes whose RNA has been degraded by incubation in the presence or absence of oxidized glutathione. Biochim Biophys Acta. 1971 Oct 14;247(2):291–303. doi: 10.1016/0005-2787(71)90677-0. [DOI] [PubMed] [Google Scholar]
  10. James D. W., Rabin B. R., Williams D. J. Role of steroid hormones in the interaction of polysomes with endoplasmic reticulum. Nature. 1969 Oct 25;224(5217):371–372. doi: 10.1038/224371a0. [DOI] [PubMed] [Google Scholar]
  11. Kosower N. S., Kosower E. M., Wertheim B., Correa W. S. Diamide, a new reagent for the intracellular oxidation of glutathione to the disulfide. Biochem Biophys Res Commun. 1969 Nov 6;37(4):593–596. doi: 10.1016/0006-291x(69)90850-x. [DOI] [PubMed] [Google Scholar]
  12. Kosower N. S., Vanderhoff G. A., Benerofe B., Hunt T., Kosower E. M. Inhibition of protein synthesis by glutathione disulfide in the presence of glutathione. Biochem Biophys Res Commun. 1971 Nov 5;45(3):816–821. doi: 10.1016/0006-291x(71)90490-6. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Lowe D., Reid E., Hallinan T. Importance of centrifugation time and homogenate concentration for isolating rat liver free polysomes: Effect of fasting. FEBS Lett. 1970 Jan 26;6(2):114–116. doi: 10.1016/0014-5793(70)80015-1. [DOI] [PubMed] [Google Scholar]
  15. Moore P. B. Reaction of N-ethyl maleimide with the ribosomes of Escherichia coli. J Mol Biol. 1971 Aug 28;60(1):169–184. doi: 10.1016/0022-2836(71)90456-6. [DOI] [PubMed] [Google Scholar]
  16. Ragland W. L., Shires T. K., Pitot H. C. Polyribosomal attachment to rat liver and hepatoma endoplasmic reticulum in vitro. A method for its study. Biochem J. 1971 Jan;121(2):271–278. doi: 10.1042/bj1210271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ranu R. S., Kaji A. Effect of sulfhydryl reagents on the ribosomes of Bacillus subtilis. J Bacteriol. 1971 Jul;107(1):53–60. doi: 10.1128/jb.107.1.53-60.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Roobol A., Rabin B. R. The binding of polysomes to smooth membranes of rat liver promoted by steroid hormones and extracts from either rough endoplasmic reticulum or from polysomes of the opposite sex. FEBS Lett. 1971 Apr 30;14(3):165–169. doi: 10.1016/0014-5793(71)80095-9. [DOI] [PubMed] [Google Scholar]
  19. Shires T. K., Narurkar L. M., Pitot H. C. Polysome interaction in vitro with microsomal membranes from rat liver. Biochem Biophys Res Commun. 1971 Dec 3;45(5):1212–1218. doi: 10.1016/0006-291x(71)90147-1. [DOI] [PubMed] [Google Scholar]
  20. Shires T. K., Narurkar L., Pitot H. C. The association in vitro of polyribosomes with ribonuclease-treated derivatives of hepatic rough endoplasmic reticulum. Characteristics of the membrane binding sites and factors influencing association. Biochem J. 1971 Nov;125(1):67–79. doi: 10.1042/bj1250067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Sunshine G. H., Williams D. J., Rabin B. R. Role for steroid hormones in the interaction of ribosomes with the endoplasmic membranes of rat liver. Nat New Biol. 1971 Mar 31;230(13):133–136. doi: 10.1038/newbio230133a0. [DOI] [PubMed] [Google Scholar]
  22. Süss R., Blobel G., Pitot H. C. Rat liver and hepatoma polysome-membrane interaction in vitro. Biochem Biophys Res Commun. 1966 May 3;23(3):299–304. doi: 10.1016/0006-291x(66)90545-6. [DOI] [PubMed] [Google Scholar]
  23. Traut R. R., Haenni A. L. The effect of sulfhydryl reagents on ribosome activity. Eur J Biochem. 1967 Jul;2(1):4–73. doi: 10.1111/j.1432-1033.1967.tb00107.x. [DOI] [PubMed] [Google Scholar]
  24. Wang J. H., Matheson A. T. The possible role of sulfhydryl groups in the dimerization of 70S ribosomes from Escherichia coli. Biochem Biophys Res Commun. 1966 Jun 13;23(5):740–744. doi: 10.1016/0006-291x(66)90463-3. [DOI] [PubMed] [Google Scholar]
  25. Williams D. J., Rabin B. R. The effects of aflatoxin B(1) and steroid hormones on polysome binding to microsomal membranes as measured by the activity of an enzyme catalysing disulphide interchange. FEBS Lett. 1969 Jul;4(2):103–107. doi: 10.1016/0014-5793(69)80207-3. [DOI] [PubMed] [Google Scholar]
  26. Zehavi-Willner T., Kosower E. M., Hunt T., Kosower N. S. Glutathione. V. The effects of the thiol-oxidizing agent diamide on initiation and translation in rabbit reticulocytes. Biochim Biophys Acta. 1971 Jan 1;228(1):245–251. [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES