Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1974 Sep;71(9):3380–3384. doi: 10.1073/pnas.71.9.3380

Highly Purified Colicin E3 Contains Immunity Protein

Karen S Jakes 1, Norton D Zinder 1
PMCID: PMC433776  PMID: 4215078

Abstract

Colicin E3, even when highly purified, still contains about one molar equivalent of a second protein, “E3 immunity protein.” The two proteins are bound together in a complex that can be dissociated only under strongly denaturing conditions, such as electrophoresis in sodium dodecyl sulfate-polyacrylamide gles or by gel filtration in 6 M guanidine·hydrochloride.

Colicin and immunity protein were separated by preparative electrophoresis in sodium dodecyl sulfate-polyacrylamide gels. The immunity protein prepared in this way was shown to be functionally and immunologically identical to immunity protein purified from colicinogenic cells by other methods. Colicin E3 that had been freed of immunity protein was much more active than complexed colicin in inhibiting protein synthesis in vitro.

Keywords: polypeptide chains, activation of colicin E3

Full text

PDF
3380

Images in this article

Selected References

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

  1. Boon T. Inactivation of ribosomes in vitro by colicin E 3 . Proc Natl Acad Sci U S A. 1971 Oct;68(10):2421–2425. doi: 10.1073/pnas.68.10.2421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bowman C. M., Dahlberg J. E., Ikemura T., Konisky J., Nomura M. Specific inactivation of 16S ribosomal RNA induced by colicin E3 in vivo. Proc Natl Acad Sci U S A. 1971 May;68(5):964–968. doi: 10.1073/pnas.68.5.964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bowman C. M., Sidikaro J., Nomura M. Specific inactivation of ribosomes by colicin E3 in vitro and mechanism of immunity in colicinogenic cells. Nat New Biol. 1971 Dec 1;234(48):133–137. doi: 10.1038/newbio234133a0. [DOI] [PubMed] [Google Scholar]
  4. Buxton R. S., Holland I. B. Colicin E2-induced DNA solubilisation in mutants of Escherichia coli deficient in endonuclease I. FEBS Lett. 1974 Feb 1;39(1):1–3. doi: 10.1016/0014-5793(74)80002-5. [DOI] [PubMed] [Google Scholar]
  5. EDELMAN G. M., OLINS D. E., GALLY J. A., ZINDER N. D. RECONSTITUTION OF IMMUNOLOGIC ACTIVITY BY INTERACTION OF POLYPEPTIDE CHAINS OF ANTIBODIES. Proc Natl Acad Sci U S A. 1963 Oct;50:753–761. doi: 10.1073/pnas.50.4.753. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. FREDERICQ P. Colicins and colicinogenic factors. Symp Soc Exp Biol. 1958;12:104–122. [PubMed] [Google Scholar]
  7. Fish W. W., Mann K. G., Tanford C. The estimation of polypeptide chain molecular weights by gel filtration in 6 M guanidine hydrochloride. J Biol Chem. 1969 Sep 25;244(18):4989–4994. [PubMed] [Google Scholar]
  8. Fromageot H. P., Zinder N. D. Growth of bacteriophage f2 in E. coli treated with rifampicin. Proc Natl Acad Sci U S A. 1968 Sep;61(1):184–191. doi: 10.1073/pnas.61.1.184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Herschman H. R., Helinski D. R. Purification and characterization of colicin E2 and colicin E3. J Biol Chem. 1967 Nov 25;242(22):5360–5368. [PubMed] [Google Scholar]
  10. Inselburg J. Colicin factor DNA: a single non-homologous region in Col E2-E3 heteroduplex molecules. Nat New Biol. 1973 Feb 21;241(112):234–237. doi: 10.1038/newbio241234a0. [DOI] [PubMed] [Google Scholar]
  11. Jakes K., Zinder N. D., Boon T. Purification and properties of colicin E3 immunity protein. J Biol Chem. 1974 Jan 25;249(2):438–444. [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Levisohn R., Konisky J., Nomura M. Interaction of colicins with bacterial cells. IV. Immunity breakdown studied with colicins Ia and Ib. J Bacteriol. 1968 Sep;96(3):811–821. doi: 10.1128/jb.96.3.811-821.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Model P., Zinder N. D. In vitro synthesis of bacteriophage f1 proteins. J Mol Biol. 1974 Feb 25;83(2):231–251. doi: 10.1016/0022-2836(74)90389-1. [DOI] [PubMed] [Google Scholar]
  15. NOMURA M. MECHANISM OF ACTION OF COLICINES. Proc Natl Acad Sci U S A. 1964 Dec;52:1514–1521. doi: 10.1073/pnas.52.6.1514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nomura M. Colicins and related bacteriocins. Annu Rev Microbiol. 1967;21:257–284. doi: 10.1146/annurev.mi.21.100167.001353. [DOI] [PubMed] [Google Scholar]
  17. Pappenheimer A. M., Jr, Gill D. M. Diphtheria. Science. 1973 Oct 26;182(4110):353–358. doi: 10.1126/science.182.4110.353. [DOI] [PubMed] [Google Scholar]
  18. Sabet S. F., Schnaitman C. A. Purification and properties of the colicin E3 receptor of Escherichia coli. J Biol Chem. 1973 Mar 10;248(5):1797–1806. [PubMed] [Google Scholar]
  19. Senior B. W., Holland I. B. Effect of colicin E3 upon the 30S ribosomal subunit of Escherichia coli. Proc Natl Acad Sci U S A. 1971 May;68(5):959–963. doi: 10.1073/pnas.68.5.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sidikaro J., Nomura M. E3 immunity substance. A protein from e3-colicinogenic cells that accounts for their immunity to colicin E3. J Biol Chem. 1974 Jan 25;249(2):445–453. [PubMed] [Google Scholar]
  21. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  22. VOGEL H. J., BONNER D. M. Acetylornithinase of Escherichia coli: partial purification and some properties. J Biol Chem. 1956 Jan;218(1):97–106. [PubMed] [Google Scholar]
  23. Weber K., Kuter D. J. Reversible denaturation of enzymes by sodium dodecyl sulfate. J Biol Chem. 1971 Jul 25;246(14):4504–4509. [PubMed] [Google Scholar]
  24. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES