Asp-51 and Asp-120 Are Important for the Transport Function of the Escherichia Coli Melibiose Carrier

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1992 May 1

PMID 1569035

Citations 7

Authors

D M Wilson

T H Wilson

Affiliations

Soon will be listed here.

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.

Citing Articles

Helical unwinding and side-chain unlocking unravel the outward open conformation of the melibiose transporter.

Wang L, Ravi V, Leblanc G, Padros E, Cladera J, Peralvarez-Marin A Sci Rep. 2016; 6:33776.

PMID: 27658476 PMC: 5034317. DOI: 10.1038/srep33776.

The Melibiose Transporter of Escherichia coli: CRITICAL CONTRIBUTION OF LYS-377 TO THE STRUCTURAL ORGANIZATION OF THE INTERACTING SUBSTRATE BINDING SITES.

Fuerst O, Lin Y, Granell M, Leblanc G, Padros E, Lorenz-Fonfria V J Biol Chem. 2015; 290(26):16261-71.

PMID: 25971963 PMC: 4481225. DOI: 10.1074/jbc.M115.642678.

Yeast nutrient transceptors provide novel insight in the functionality of membrane transporters.

Schothorst J, Kankipati H, Conrad M, Samyn D, Van Zeebroeck G, Popova Y Curr Genet. 2013; 59(4):197-206.

PMID: 24114446 PMC: 3824880. DOI: 10.1007/s00294-013-0413-y.

Structural insights into the activation mechanism of melibiose permease by sodium binding.

Granell M, Leon X, Leblanc G, Padros E, Lorenz-Fonfria V Proc Natl Acad Sci U S A. 2010; 107(51):22078-83.

PMID: 21135207 PMC: 3009771. DOI: 10.1073/pnas.1008649107.

Alteration of sugar-induced conformational changes of the melibiose permease by mutating Arg141 in loop 4-5.

Leon X, Leblanc G, Padros E Biophys J. 2009; 96(12):4877-86.

PMID: 19527646 PMC: 2712053. DOI: 10.1016/j.bpj.2009.03.025.

References

Tsuchiya T, Wilson T . Cation-sugar cotransport in the melibiose transport system of Escherichia coli. Membr Biochem. 1978; 2(1):63-79. DOI: 10.3109/09687687809063858. View

Ryden S, ISAKSSON L . 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. View

Kleina L, MASSON J, Normanly J, Abelson J, Miller J . Construction of Escherichia coli amber suppressor tRNA genes. II. Synthesis of additional tRNA genes and improvement of suppressor efficiency. J Mol Biol. 1990; 213(4):705-17. DOI: 10.1016/S0022-2836(05)80257-8. View

Botfield M, Wilson T . 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; 264(20):11649-52. View

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; 162(3):933-7. PMC: 215865. DOI: 10.1128/jb.162.3.933-937.1985. View

Botfield M, Wilson T . Mutations that simultaneously alter both sugar and cation specificity in the melibiose carrier of Escherichia coli. J Biol Chem. 1988; 263(26):12909-15. View

Lolkema J, Puttner I, Kaback H . Site-directed mutagenesis of Pro327 in the lac permease of Escherichia coli. Biochemistry. 1988; 27(22):8307-10. DOI: 10.1021/bi00422a003. View

Wilson D, Wilson T . Cation specificity for sugar substrates of the melibiose carrier in Escherichia coli. Biochim Biophys Acta. 1987; 904(2):191-200. DOI: 10.1016/0005-2736(87)90368-3. View

Hanatani M, Yazyu H, 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; 259(3):1807-12. View

10.

Tsuchiya T, Oho M . Lithium ion-sugar cotransport via the melibiose transport system in Escherichia coli. Measurement of Li+ transport and specificity. J Biol Chem. 1983; 258(21):12765-7. View

11.

Niiya S, Yamasaki K, Wilson T, Tsuchiya T . Altered cation coupling to melibiose transport in mutants of Escherichia coli. J Biol Chem. 1982; 257(15):8902-6. View

12.

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

13.

Schreier P, Cortese R . A fast and simple method for sequencing DNA cloned in the single-stranded bacteriophage M13. J Mol Biol. 1979; 129(1):169-72. DOI: 10.1016/0022-2836(79)90068-8. View

14.

Pourcher T, Bassilana M, Sarkar H, Kaback H, Leblanc G . The melibiose/Na+ symporter of Escherichia coli: kinetic and molecular properties. Philos Trans R Soc Lond B Biol Sci. 1990; 326(1236):411-23. DOI: 10.1098/rstb.1990.0021. View

15.

Pourcher T, Sarkar H, Bassilana M, Kaback H, 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; 87(1):468-72. PMC: 53285. DOI: 10.1073/pnas.87.1.468. View

16.

Botfield M, Wilson T . Carboxyl-terminal truncations of the melibiose carrier of Escherichia coli. J Biol Chem. 1989; 264(20):11643-8. View

17.

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; 263(28):14276-80. View

18.

Taylor J, 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; 13(24):8749-64. PMC: 318949. DOI: 10.1093/nar/13.24.8749. View

19.

Taylor J, Ott J, Eckstein F . The rapid generation of oligonucleotide-directed mutations at high frequency using phosphorothioate-modified DNA. Nucleic Acids Res. 1985; 13(24):8765-85. PMC: 318950. DOI: 10.1093/nar/13.24.8765. View

20.

Eggertsson G, Soll D . Transfer ribonucleic acid-mediated suppression of termination codons in Escherichia coli. Microbiol Rev. 1988; 52(3):354-74. PMC: 373150. DOI: 10.1128/mr.52.3.354-374.1988. View