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.
The first step in the transport of cyanocobalamin (CN-B(12)) by cells of Escherichia coli was shown previously to consist of binding of the B(12) to specific receptor sites located on the outer membrane of the cell envelope. In this paper, evidence is presented that these B(12) receptor sites also function as the receptors for the E colicins, and that there is competition between B(12) and the E colicins for occupancy of these sites. The cell strains used were E. coli KBT001, a methionine/B(12) auxotroph, and B(12) transport mutants derived from strain KBT001. Colicins E1 and E3 inhibited binding of B(12) to the outer membrane B(12) receptor sites, and CN-B(12) protected cells against these colicins. Half-maximal protection was given by CN-B(12) concentrations in the range of 1 to 6 nM, depending upon the colicin concentration used. Colicin E1 competitively inhibited the binding of (57)Co-labeled CN-B(12) to isolated outer membrane particles. Functional colicin E receptor sites were found in cell envelopes from cells of only those strains that possessed intact B(12) receptors. Colicin K did not inhibit the binding of B(12) to the outer membrane receptor sites, and no evidence was found for any identity between the B(12) and colicin K receptors. However, both colicin K and colicin E1 inhibited the secondary phase of B(12) transport, which is believed to consist of the energy-coupled movement of B(12) across the inner membrane.
D000596:Amino Acids / Q000378:metabolism; D001426:Bacterial Proteins / Q000096:biosynthesis; D001665:Binding Sites; D001693:Biological Transport, Active; D002247:Carbon Isotopes; D002473:Cell Wall / Q000378:metabolism; D003036:Cobalt Isotopes; D003087:Colicins / Q000378:metabolism* / Q000494:pharmacology; D004926:Escherichia coli / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008715:Methionine / Q000378:metabolism; D009154:Mutation; D011955:Receptors, Drug; D012044:Regression Analysis; D013045:Species Specificity; D013237:Stereoisomerism; D014805:Vitamin B 12 / Q000378:metabolism*
D000596:Amino Acids; D001426:Bacterial Proteins; D002247:Carbon Isotopes; D003036:Cobalt Isotopes; D003087:Colicins; D011955:Receptors, Drug; D008715:Methionine; D007930:Leucine; D008239:Lysine; D014805:Vitamin B 12