Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1984 Oct;160(1):385–390. doi: 10.1128/jb.160.1.385-390.1984

Effect of chloride and glutamate ions on in vitro protein synthesis by the moderate halophile Vibrio costicola.

M Kamekura, D J Kushner
PMCID: PMC214729  PMID: 6148335

Abstract

Vibrio costicola grown in the presence of different NaCl concentrations contains cell-associated Na+ and K+ ions whose sum is equal to or greater than the external Na+ concentration. In the presence of 0.5 M NaCl, virtually no in vitro protein is synthesized in extracts of cells grown in 1.0 M NaCl. However, we report here that active in vitro protein synthesis occurred in 0.6 M or higher concentrations of Na2SO4, sodium formate, sodium acetate, sodium aspartate, or sodium glutamate, whereas 0.6 M NaF, NaCl, or NaBr completely inhibited protein synthesis as measured by polyuridylic acid-directed incorporation of [14C]phenylalanine. Sodium glutamate, sodium aspartate, and betaine (0.3 M) counteracted the inhibitory action of 0.6 M NaCl. The cell-associated Cl- concentration was 0.22 mol/kg in cells grown in 1.0 M NaCl. Of this, the free intracellular Cl- concentration was only 0.02 mol/kg. Cells contained 0.11 mol of glutamate per kg and small concentrations of other amino acids. All of the negative counterions for cell-associated Na+ and K+ have not yet been determined. In vitro protein synthesis by Escherichia coli was inhibited by sodium glutamate. Hybridization experiments with ribosomes and the soluble (S-100) fractions from extracts of E. coli and V. costicola showed that the glutamate-sensitive fraction was found in the soluble, not the ribosomal, part of the system. The phenylalanyl-tRNA synthetase of V. costicola was not inhibited by 0.5 M or higher concentrations of NaCl; it was slightly more sensitive to high concentrations of sodium glutamate. Therefore, this enzyme was not responsible for the salt response of the V. costicola in vitro protein-synthesizing system.

Full text

PDF
389

Selected References

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

  1. Bayley S. T., Griffiths E. A cell-free amino acid incorporating system from an extremely halophilic bacterium. Biochemistry. 1968 Jun;7(6):2249–2256. doi: 10.1021/bi00846a030. [DOI] [PubMed] [Google Scholar]
  2. Bengis-Garber C., Gromet-Elhanan Z. Purification of the energy-transducing adenosine triphosphatase complex from Rhodospirillum rubrum. Biochemistry. 1979 Aug 7;18(16):3577–3581. doi: 10.1021/bi00583a022. [DOI] [PubMed] [Google Scholar]
  3. CHRISTIAN J. H., WALTHO J. A. Solute concentrations within cells of halophilic and non-halophilic bacteria. Biochim Biophys Acta. 1962 Dec 17;65:506–508. doi: 10.1016/0006-3002(62)90453-5. [DOI] [PubMed] [Google Scholar]
  4. Coleman G. Nature of the major inorganic ions concentrated during growth of Bacillus amyloliquefaciens. J Gen Microbiol. 1974 Oct;84(2):297–302. doi: 10.1099/00221287-84-2-297. [DOI] [PubMed] [Google Scholar]
  5. Di Segni G., Rosen H., Kaempfer R. Competition between alpha- and beta-globin messenger ribonucleic acids for eucaryotic initiation factor 2. Biochemistry. 1979 Jun 26;18(13):2847–2854. doi: 10.1021/bi00580a027. [DOI] [PubMed] [Google Scholar]
  6. Furano A. V. A very rapid method for washing large numbers of precipitates of proteins and nucleic acids. Anal Biochem. 1971 Oct;43(2):639–640. doi: 10.1016/0003-2697(71)90300-9. [DOI] [PubMed] [Google Scholar]
  7. Hipkiss A. R., Armstrong D. W., Kushner D. J. Protein turnover in a moderately halophilic bacterium. Can J Microbiol. 1980 Feb;26(2):196–203. doi: 10.1139/m80-030. [DOI] [PubMed] [Google Scholar]
  8. Kushner D. J., Hamaide F., MacLeod R. A. Development of salt-resistant active transport in a moderately halophilic bacterium. J Bacteriol. 1983 Mar;153(3):1163–1171. doi: 10.1128/jb.153.3.1163-1171.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Masui M., Wada S. Intracellular concentrations of Na+, K+, and cl minus of a moderately halophilic bacterium. Can J Microbiol. 1973 Oct;19(10):1181–1186. doi: 10.1139/m73-191. [DOI] [PubMed] [Google Scholar]
  11. SCHULTZ S. G., WILSON N. L., EPSTEIN W. Cation transport in Escherichia coli. II. Intracellular chloride concentration. J Gen Physiol. 1962 Sep;46:159–166. doi: 10.1085/jgp.46.1.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Shindler D. B., Wydro R. M., Kushner D. J. Cell-bound cations of the moderately halophilic bacterium Vibrio costicola. J Bacteriol. 1977 May;130(2):698–703. doi: 10.1128/jb.130.2.698-703.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Tuite M. F., Plesset J., Moldave K., McLaughlin C. S. Faithful and efficient translation of homologous and heterologous mRNAs in an mRNA-dependent cell-free system from Saccharomyces cerevisiae. J Biol Chem. 1980 Sep 25;255(18):8761–8766. [PubMed] [Google Scholar]
  14. Weber L. A., Hickey E. D., Maroney P. A., Baglioni C. Inhibition of protein synthesis by Cl-. J Biol Chem. 1977 Jun 10;252(11):4007–4010. [PubMed] [Google Scholar]
  15. Wydro R. M., Madira W., Hiramatsu T., Kogut M., Kushner D. J. Salt-sensitive in vitro protein synthesis by a moderately halophilic bacterium. Nature. 1977 Oct 27;269(5631):824–825. doi: 10.1038/269824a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES