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. 1973 Aug;115(2):506–513. doi: 10.1128/jb.115.2.506-513.1973

Transport of Vitamin B12 in Escherichia coli: Common Receptor Sites for Vitamin B12 and the E Colicins on the Outer Membrane of the Cell Envelope

Donna R Di Masi 1, John C White 1, Carl A Schnaitman 1, Clive Bradbeer 1
PMCID: PMC246277  PMID: 4579869

Abstract

The first step in the transport of cyanocobalamin (CN-B12) by cells of Escherichia coli was shown previously to consist of binding of the B12 to specific receptor sites located on the outer membrane of the cell envelope. In this paper, evidence is presented that these B12 receptor sites also function as the receptors for the E colicins, and that there is competition between B12 and the E colicins for occupancy of these sites. The cell strains used were E. coli KBT001, a methionine/B12 auxotroph, and B12 transport mutants derived from strain KBT001. Colicins E1 and E3 inhibited binding of B12 to the outer membrane B12 receptor sites, and CN-B12 protected cells against these colicins. Half-maximal protection was given by CN-B12 concentrations in the range of 1 to 6 nM, depending upon the colicin concentration used. Colicin E1 competitively inhibited the binding of 57Co-labeled CN-B12 to isolated outer membrane particles. Functional colicin E receptor sites were found in cell envelopes from cells of only those strains that possessed intact B12 receptors. Colicin K did not inhibit the binding of B12 to the outer membrane receptor sites, and no evidence was found for any identity between the B12 and colicin K receptors. However, both colicin K and colicin E1 inhibited the secondary phase of B12 transport, which is believed to consist of the energy-coupled movement of B12 across the inner membrane.

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Selected References

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

  1. Bhattacharyya P., Wendt L., Whitney E., Silver S. Colicin-tolerant mutants of Escherichia coli: resistance of membranes to colicin E1. Science. 1970 May 22;168(3934):998–1000. doi: 10.1126/science.168.3934.998. [DOI] [PubMed] [Google Scholar]
  2. Buxton R. S. Genetic analysis of Escherichia coli K12 mutants resistant to bacteriophage BF23 and the E-group colicins. Mol Gen Genet. 1971;113(2):154–156. doi: 10.1007/BF00333188. [DOI] [PubMed] [Google Scholar]
  3. DAVIS B. D., MINGIOLI E. S. Mutants of Escherichia coli requiring methionine or vitamin B12. J Bacteriol. 1950 Jul;60(1):17–28. doi: 10.1128/jb.60.1.17-28.1950. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Di Girolamo P. M., Bradbeer C. Transport of vitamin B 12 in Escherichia coli. J Bacteriol. 1971 Jun;106(3):745–750. doi: 10.1128/jb.106.3.745-750.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Di Girolamo P. M., Kadner R. J., Bradbeer C. Isolation of vitamin B 12 transport mutants of Escherichia coli. J Bacteriol. 1971 Jun;106(3):751–757. doi: 10.1128/jb.106.3.751-757.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. FREDERICQ P. Recherches sur les colicines du groupe E. C R Seances Soc Biol Fil. 1950 Feb;144(3-4):297–299. [PubMed] [Google Scholar]
  7. Fields K. L., Luria S. E. Effects of colicins E1 and K on transport systems. J Bacteriol. 1969 Jan;97(1):57–63. doi: 10.1128/jb.97.1.57-63.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Kadner R. J., Liggins G. L. Transport of vitamin B12 in Escherichia coli: genetic studies. J Bacteriol. 1973 Aug;115(2):514–521. doi: 10.1128/jb.115.2.514-521.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Sabet S. F., Schnaitman C. A. Localization and solubilization of colicin receptors. J Bacteriol. 1971 Oct;108(1):422–430. doi: 10.1128/jb.108.1.422-430.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Schnaitman C. A. Protein composition of the cell wall and cytoplasmic membrane of Escherichia coli. J Bacteriol. 1970 Nov;104(2):890–901. doi: 10.1128/jb.104.2.890-901.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Schnaitman C. A. Solubilization of the cytoplasmic membrane of Escherichia coli by Triton X-100. J Bacteriol. 1971 Oct;108(1):545–552. doi: 10.1128/jb.108.1.545-552.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. WADDELL W. J. A simple ultraviolet spectrophotometric method for the determination of protein. J Lab Clin Med. 1956 Aug;48(2):311–314. [PubMed] [Google Scholar]
  15. White J. C., DiGirolamo P. M., Fu M. L., Preston Y. A., Bradbeer C. Transport of vitamin B 12 in Escherichia coli. Location and properties of the initial B 12 -binding site. J Biol Chem. 1973 Jun 10;248(11):3978–3986. [PubMed] [Google Scholar]

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