Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1985 Jun;162(3):933–937. doi: 10.1128/jb.162.3.933-937.1985

Alteration in cation specificity of the melibiose transport carrier of Escherichia coli due to replacement of proline 122 with serine.

H Yazyu, S Shiota, M Futai, T Tsuchiya
PMCID: PMC215865  PMID: 2987195

Abstract

The structural genes (melB) for the melibiose carrier of five mutants of Escherichia coli showing altered cation specificity for melibiose transport were cloned. The mutations were mapped in a 248-base-pair DNA fragment by a recombinational assay by using the mutants transformed with hybrid plasmids carrying various portions of the wild-type melB gene. The nucleotide sequences of the corresponding DNA fragments derived from mutated melB genes were determined, and the amino acid sequences of the carrier were deduced. Proline 122 was replaced with serine in the melibiose carrier of all five mutants (which were isolated independently). We conclude that this amino acid replacement caused the alteration in cation specificity (loss of coupling to H+) of the melibiose carrier.

Full text

PDF
934

Images in this article

936Image
on p.936

Selected References

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

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Chothia C., Levitt M., Richardson D. Structure of proteins: packing of alpha-helices and pleated sheets. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4130–4134. doi: 10.1073/pnas.74.10.4130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chou P. Y., Fasman G. D. Empirical predictions of protein conformation. Annu Rev Biochem. 1978;47:251–276. doi: 10.1146/annurev.bi.47.070178.001343. [DOI] [PubMed] [Google Scholar]
  5. Coulondre C., Miller J. H. Genetic studies of the lac repressor. IV. Mutagenic specificity in the lacI gene of Escherichia coli. J Mol Biol. 1977 Dec 15;117(3):577–606. doi: 10.1016/0022-2836(77)90059-6. [DOI] [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. Kyte J., Doolittle R. F. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. doi: 10.1016/0022-2836(82)90515-0. [DOI] [PubMed] [Google Scholar]
  8. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  9. Levitt M. Effect of proline residues on protein folding. J Mol Biol. 1981 Jan 5;145(1):251–263. doi: 10.1016/0022-2836(81)90342-9. [DOI] [PubMed] [Google Scholar]
  10. Lopilato J., Tsuchiya T., Wilson T. H. Role of Na+ and Li+ in thiomethylgalactoside transport by the melibiose transport system of Escherichia coli. J Bacteriol. 1978 Apr;134(1):147–156. doi: 10.1128/jb.134.1.147-156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  12. Molholt B., Doskocil J. Increased transformation frequency in E. coli. Biochem Biophys Res Commun. 1978 May 30;82(2):477–483. doi: 10.1016/0006-291x(78)90899-9. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Schendel P. F., Wells R. D. The synthesis and purification of (gamma-32P)-adenosine triphosphate with high specific activity. J Biol Chem. 1973 Dec 10;248(23):8319–8321. [PubMed] [Google Scholar]
  15. 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]
  16. Tanaka S., Lerner S. A., Lin E. C. Replacement of a phosphoenolpyruvate-dependent phosphotransferase by a nicotinamide adenine dinucleotide-linked dehydrogenase for the utilization of mannitol. J Bacteriol. 1967 Feb;93(2):642–648. doi: 10.1128/jb.93.2.642-648.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Trumble W. R., Viitanen P. V., Sarkar H. K., Poonian M. S., Kaback H. R. Site-directed mutagenesis of cys148 in the lac carrier protein of Escherichia coli. Biochem Biophys Res Commun. 1984 Mar 30;119(3):860–867. doi: 10.1016/0006-291x(84)90853-2. [DOI] [PubMed] [Google Scholar]
  18. Tsuchiya T., Lopilato J., Wilson T. H. Effect of lithium ion on melibiose transport in Escherichia coli. J Membr Biol. 1978 Jul 21;42(1):45–59. doi: 10.1007/BF01870393. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. 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]

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

RESOURCES