Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1978 Oct;136(1):168–174. doi: 10.1128/jb.136.1.168-174.1978

Genetic separation of high- and low-affinity transport systems for branched-chain amino acids in Escherichia coli K-12.

J J Anderson, D L Oxender
PMCID: PMC218646  PMID: 361686

Abstract

The Escherichia coli K-12 mutant strain AE4107 (livH::Mu) is defective in the high-affinity binding protein-mediated uptake system for L-leucine, L-valine, and L-isoleucine (LIV-I). We have used this strain to produce mutations in the residual LIV-II membrane-bound branched-chain amino acid uptake system. Mutants selected for their inability to utilize exogenous L-leucine were found to be defective in the LIV-II system and fell into two classes. One class, represented by strain AE410709 (livP9), showed a complete loss of saturable uptake for L-leucine, L-valine, and L-isoleucine up to 50 muM, and a second class, represented by strain AE4017012 (liv-12), showed a residual component of saturable leucine uptake with increased Km. These mutations, livP9 and liv-12, were closely linked and mapped in the 74 to 78 min region of the E. coli genetic map. Strains constructed so that they lacked both LIV-I and LIV-II transport systems excreted leucine. Strains of the genotype livH+ livP were found to have normal high-affinity binding protein-mediated transport (LIV-I and leucine specific), whereas the low-affinity (LIV-II) transport was completely missing. We concluded from these studies that the high-affinity binding protein-mediated transport systems (LIV-I and leucine specific) can operate independently of the membrane-bound LIV-II system.

Full text

PDF
168

Selected References

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

  1. Anderson J. J., Oxender D. L. Escherichia coli transport mutants lacking binding protein and other components of the branched-chain amino acid transport systems. J Bacteriol. 1977 Apr;130(1):384–392. doi: 10.1128/jb.130.1.384-392.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson J. J., Quay S. C., Oxender D. L. Mapping of two loci affecting the regulation of branched-chain amino acid transport in Escherichia coli K-12. J Bacteriol. 1976 Apr;126(1):80–90. doi: 10.1128/jb.126.1.80-90.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark D., Cronan J. E., Jr Further mapping of several membrane lipid biosynthetic genes (fabC, fabB, gpsA, plsB) of Escherichia coli. J Bacteriol. 1977 Nov;132(2):549–554. doi: 10.1128/jb.132.2.549-554.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cornish-Bowden A., Eisenthal R. Statistical considerations in the estimation of enzyme kinetic parameters by the direct linear plot andother methods. Biochem J. 1974 Jun;139(3):721–730. doi: 10.1042/bj1390721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Eisenthal R., Cornish-Bowden A. The direct linear plot. A new graphical procedure for estimating enzyme kinetic parameters. Biochem J. 1974 Jun;139(3):715–720. doi: 10.1042/bj1390715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Furlong C. E., Weiner J. H. Purification of a leucine-specific binding protein from Escherichia coli. Biochem Biophys Res Commun. 1970 Mar 27;38(6):1076–1083. doi: 10.1016/0006-291x(70)90349-9. [DOI] [PubMed] [Google Scholar]
  8. Guardiola J., De Felice M., Klopotowski T., Iaccarino M. Multiplicity of isoleucine, leucine, and valine transport systems in Escherichia coli K-12. J Bacteriol. 1974 Feb;117(2):382–392. doi: 10.1128/jb.117.2.382-392.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Koch A. L. Turbidity measurements of bacterial cultures in some available commercial instruments. Anal Biochem. 1970 Nov;38(1):252–259. doi: 10.1016/0003-2697(70)90174-0. [DOI] [PubMed] [Google Scholar]
  10. Neidhardt F. C., Bloch P. L., Smith D. F. Culture medium for enterobacteria. J Bacteriol. 1974 Sep;119(3):736–747. doi: 10.1128/jb.119.3.736-747.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Quay S. C., Oxender D. L. Regulation of branched-chain amino acid transport in Escherichia coli. J Bacteriol. 1976 Sep;127(3):1225–1238. doi: 10.1128/jb.127.3.1225-1238.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Rahmanian M., Claus D. R., Oxender D. L. Multiplicity of leucine transport systems in Escherichia coli K-12. J Bacteriol. 1973 Dec;116(3):1258–1266. doi: 10.1128/jb.116.3.1258-1266.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Robbins A. R., Rotman B. Evidence for binding protein-independent substrate translocation by the methylgalactoside transport system of Escherichia coli K12. Proc Natl Acad Sci U S A. 1975 Feb;72(2):423–427. doi: 10.1073/pnas.72.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Rosen B. P. Basic amino acid transport in Escherichia coli: properties of canavanine-resistant mutants. J Bacteriol. 1973 Nov;116(2):627–635. doi: 10.1128/jb.116.2.627-635.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Shifrin S., Ames B. N., FerroLuzzi-Ames G. Effect of the alpha-hydrazino analogue of histidine on histidine transport and arginine biosynthesis. J Biol Chem. 1966 Jul 25;241(14):3424–3429. [PubMed] [Google Scholar]
  16. Wood J. M. Leucine transport in Escherichia coli. The resolution of multiple transport systems and their coupling to metabolic energy. J Biol Chem. 1975 Jun 25;250(12):4477–4485. [PubMed] [Google Scholar]
  17. Yamato I., Anraku Y. Transport of sugars and amino acids in bacteria. XVIII. Properties of an isoleucine carrier in the cytoplasmic membrane vesicles of Escherichia coli. J Biochem. 1977 May;81(5):1517–1523. [PubMed] [Google Scholar]

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

RESOURCES