Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 May;174(9):3083–3086. doi: 10.1128/jb.174.9.3083-3086.1992

Asp-51 and Asp-120 are important for the transport function of the Escherichia coli melibiose carrier.

D M Wilson 1, T H Wilson 1
PMCID: PMC205965  PMID: 1569035

Abstract

Asp-51 and Asp-120 of the Escherichia coli melibiose carrier on plasmid pKKMB were separately replaced by amber codons and transformed into eight amber suppressor strains, producing eight amino acid substitutions for each site. Glu-51 and Glu-120 were the only replacements in the carrier that allowed the cells to ferment melibiose and that showed transport of melibiose against a concentration gradient. Revertants to Glu-51 and Glu-120 show less activity than the wild type. The Asp-51 position is more crucial for Na(+)-stimulated melibiose accumulation than is the Asp-120 site.

Full text

PDF
3083

Selected References

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

  1. Botfield M. C., Wilson T. H. Carboxyl-terminal truncations of the melibiose carrier of Escherichia coli. J Biol Chem. 1989 Jul 15;264(20):11643–11648. [PubMed] [Google Scholar]
  2. Botfield M. C., Wilson T. H. Mutations that simultaneously alter both sugar and cation specificity in the melibiose carrier of Escherichia coli. J Biol Chem. 1988 Sep 15;263(26):12909–12915. [PubMed] [Google Scholar]
  3. Botfield M. C., Wilson T. H. Peptide-specific antibody for the melibiose carrier of Escherichia coli localizes the carboxyl terminus to the cytoplasmic face of the membrane. J Biol Chem. 1989 Jul 15;264(20):11649–11652. [PubMed] [Google Scholar]
  4. Certa U., Bannwarth W., Stüber D., Gentz R., Lanzer M., Le Grice S., Guillot F., Wendler I., Hunsmann G., Bujard H. Subregions of a conserved part of the HIV gp41 transmembrane protein are differentially recognized by antibodies of infected individuals. EMBO J. 1986 Nov;5(11):3051–3056. doi: 10.1002/j.1460-2075.1986.tb04605.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eggertsson G., Söll D. Transfer ribonucleic acid-mediated suppression of termination codons in Escherichia coli. Microbiol Rev. 1988 Sep;52(3):354–374. doi: 10.1128/mr.52.3.354-374.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hanatani M., Yazyu H., Shiota-Niiya S., Moriyama Y., Kanazawa H., Futai M., Tsuchiya T. Physical and genetic characterization of the melibiose operon and identification of the gene products in Escherichia coli. J Biol Chem. 1984 Feb 10;259(3):1807–1812. [PubMed] [Google Scholar]
  7. Kawakami T., Akizawa Y., Ishikawa T., Shimamoto T., Tsuda M., Tsuchiya T. Amino acid substitutions and alteration in cation specificity in the melibiose carrier of Escherichia coli. J Biol Chem. 1988 Oct 5;263(28):14276–14280. [PubMed] [Google Scholar]
  8. Kleina L. G., Masson J. M., Normanly J., Abelson J., Miller J. H. Construction of Escherichia coli amber suppressor tRNA genes. II. Synthesis of additional tRNA genes and improvement of suppressor efficiency. J Mol Biol. 1990 Jun 20;213(4):705–717. doi: 10.1016/S0022-2836(05)80257-8. [DOI] [PubMed] [Google Scholar]
  9. Kraft R., Tardiff J., Krauter K. S., Leinwand L. A. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques. 1988 Jun;6(6):544-6, 549. [PubMed] [Google Scholar]
  10. Lolkema J. S., Püttner I. B., Kaback H. R. Site-directed mutagenesis of Pro327 in the lac permease of Escherichia coli. Biochemistry. 1988 Nov 1;27(22):8307–8310. doi: 10.1021/bi00422a003. [DOI] [PubMed] [Google Scholar]
  11. Niiya S., Yamasaki K., Wilson T. H., Tsuchiya T. Altered cation coupling to melibiose transport in mutants of Escherichia coli. J Biol Chem. 1982 Aug 10;257(15):8902–8906. [PubMed] [Google Scholar]
  12. Pourcher T., Bassilana M., Sarkar H. K., Kaback H. R., Leblanc G. The melibiose/Na+ symporter of Escherichia coli: kinetic and molecular properties. Philos Trans R Soc Lond B Biol Sci. 1990 Jan 30;326(1236):411–423. doi: 10.1098/rstb.1990.0021. [DOI] [PubMed] [Google Scholar]
  13. Pourcher T., Deckert M., Bassilana M., Leblanc G. Melibiose permease of Escherichia coli: mutation of aspartic acid 55 in putative helix II abolishes activation of sugar binding by Na+ ions. Biochem Biophys Res Commun. 1991 Aug 15;178(3):1176–1181. doi: 10.1016/0006-291x(91)91016-6. [DOI] [PubMed] [Google Scholar]
  14. Pourcher T., Sarkar H. K., Bassilana M., Kaback H. R., Leblanc G. Histidine-94 is the only important histidine residue in the melibiose permease of Escherichia coli. Proc Natl Acad Sci U S A. 1990 Jan;87(1):468–472. doi: 10.1073/pnas.87.1.468. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Rydén S. M., Isaksson L. A. A temperature-sensitive mutant of Escherichia coli that shows enhanced misreading of UAG/A and increased efficiency for some tRNA nonsense suppressors. Mol Gen Genet. 1984;193(1):38–45. doi: 10.1007/BF00327411. [DOI] [PubMed] [Google Scholar]
  16. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schreier P. H., Cortese R. A fast and simple method for sequencing DNA cloned in the single-stranded bacteriophage M13. J Mol Biol. 1979 Mar 25;129(1):169–172. doi: 10.1016/0022-2836(79)90068-8. [DOI] [PubMed] [Google Scholar]
  18. Shiota S., Yamane Y., Futai M., Tsuchiya T. Escherichia coli mutants possessing an Li+-resistant melibiose carrier. J Bacteriol. 1985 Apr;162(1):106–109. doi: 10.1128/jb.162.1.106-109.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shiota S., Yazyu H., Tsuchiya T. Escherichia coli mutants with altered cation recognition by the melibiose carrier. J Bacteriol. 1984 Oct;160(1):445–447. doi: 10.1128/jb.160.1.445-447.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Taylor J. W., Ott J., Eckstein F. The rapid generation of oligonucleotide-directed mutations at high frequency using phosphorothioate-modified DNA. Nucleic Acids Res. 1985 Dec 20;13(24):8765–8785. doi: 10.1093/nar/13.24.8765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Taylor J. W., Schmidt W., Cosstick R., Okruszek A., Eckstein F. The use of phosphorothioate-modified DNA in restriction enzyme reactions to prepare nicked DNA. Nucleic Acids Res. 1985 Dec 20;13(24):8749–8764. doi: 10.1093/nar/13.24.8749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Tsuchiya T., Oho M., Shiota-Niiya S. Lithium ion-sugar cotransport via the melibiose transport system in Escherichia coli. Measurement of Li+ transport and specificity. J Biol Chem. 1983 Nov 10;258(21):12765–12767. [PubMed] [Google Scholar]
  23. Tsuchiya T., Wilson T. H. Cation-sugar cotransport in the melibiose transport system of Escherichia coli. Membr Biochem. 1978;2(1):63–79. doi: 10.3109/09687687809063858. [DOI] [PubMed] [Google Scholar]
  24. Wilson D. M., Wilson T. H. Cation specificity for sugar substrates of the melibiose carrier in Escherichia coli. Biochim Biophys Acta. 1987 Nov 13;904(2):191–200. doi: 10.1016/0005-2736(87)90368-3. [DOI] [PubMed] [Google Scholar]
  25. Yazyu H., Shiota-Niiya S., Shimamoto T., Kanazawa H., Futai M., Tsuchiya T. Nucleotide sequence of the melB gene and characteristics of deduced amino acid sequence of the melibiose carrier in Escherichia coli. J Biol Chem. 1984 Apr 10;259(7):4320–4326. [PubMed] [Google Scholar]
  26. Yazyu H., Shiota S., Futai M., Tsuchiya T. Alteration in cation specificity of the melibiose transport carrier of Escherichia coli due to replacement of proline 122 with serine. J Bacteriol. 1985 Jun;162(3):933–937. doi: 10.1128/jb.162.3.933-937.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES