Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Dec;169(12):5648–5652. doi: 10.1128/jb.169.12.5648-5652.1987

The blue copper protein gene of Alcaligenes faecalis S-6 directs secretion of blue copper protein from Escherichia coli cells.

K Yamamoto 1, T Uozumi 1, T Beppu 1
PMCID: PMC214024  PMID: 2824441

Abstract

The gene encoding a blue copper protein (a member of the pseudoazurins) of 123 amino acid residues, containing a single type I Cu2+ ion, was cloned from Alcaligenes faecalis S-6. The nucleotide sequence of the coding region, as well as the 5'- and 3'-flanking regions, was determined. The deduced amino acid sequence after Glu-24 coincided with the reported sequence of the blue protein, and its NH2-terminal sequence of 23 residues resembled a typical signal peptide. The cloned gene was expressed under the control of the tac promoter in Escherichia coli, and the correctly processed blue protein was secreted into the periplasm. The blue protein produced in E. coli possessed the activity to transfer electrons to the copper-containing nitrite reductase of A. faecalis S-6 in vitro.

Full text

PDF
5651

Images in this article

5651Image
on p.5651
5651Image
on p.5651

Selected References

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

  1. Ambler R. P., Tobari J. The primary structures of Pseudomonas AM1 amicyanin and pseudoazurin. Two new sequence classes of blue copper proteins. Biochem J. 1985 Dec 1;232(2):451–457. doi: 10.1042/bj2320451. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Canters G. W. The azurin gene from Pseudomonas aeruginosa codes for a pre-protein with a signal peptide. Cloning and sequencing of the azurin gene. FEBS Lett. 1987 Feb 9;212(1):168–172. doi: 10.1016/0014-5793(87)81579-x. [DOI] [PubMed] [Google Scholar]
  3. Cornelis P., Digneffe C., Willemot K. Cloning and expression of a Bacillus coagulans amylase gene in Escherichia coli. Mol Gen Genet. 1982;186(4):507–511. doi: 10.1007/BF00337957. [DOI] [PubMed] [Google Scholar]
  4. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  5. Hormel S., Adman E., Walsh K. A., Beppu T., Titani K. The amino acid sequence of the blue copper protein of Alcaligenes faecalis. FEBS Lett. 1986 Mar 3;197(1-2):301–304. doi: 10.1016/0014-5793(86)80346-5. [DOI] [PubMed] [Google Scholar]
  6. Hunt T. P., Magasanik B. Transcription of glnA by purified Escherichia coli components: core RNA polymerase and the products of glnF, glnG, and glnL. Proc Natl Acad Sci U S A. 1985 Dec;82(24):8453–8457. doi: 10.1073/pnas.82.24.8453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Husain M., Davidson V. L. An inducible periplasmic blue copper protein from Paracoccus denitrificans. Purification, properties, and physiological role. J Biol Chem. 1985 Nov 25;260(27):14626–14629. [PubMed] [Google Scholar]
  8. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kakutani T., Watanabe H., Arima K., Beppu T. A blue protein as an inactivating factor for nitrite reductase from Alcaligenes faecalis strain S-6. J Biochem. 1981 Feb;89(2):463–472. doi: 10.1093/oxfordjournals.jbchem.a133221. [DOI] [PubMed] [Google Scholar]
  10. Kakutani T., Watanabe H., Arima K., Beppu T. Purification and properties of a copper-containing nitrite reductase from a denitrifying bacterium, Alcaligenes faecalis strain S-6. J Biochem. 1981 Feb;89(2):453–461. doi: 10.1093/oxfordjournals.jbchem.a133220. [DOI] [PubMed] [Google Scholar]
  11. Liu M. Y., Liu M. C., Payne W. J., Legall J. Properties and electron transfer specificity of copper proteins from the denitrifier "Achromobacter cycloclastes". J Bacteriol. 1986 May;166(2):604–608. doi: 10.1128/jb.166.2.604-608.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Norgard M. V., Keem K., Monahan J. J. Factors affecting the transformation of Escherichia coli strain chi1776 by pBR322 plasmid DNA. Gene. 1978 Jul;3(4):279–292. doi: 10.1016/0378-1119(78)90038-0. [DOI] [PubMed] [Google Scholar]
  13. Norris G. E., Anderson B. F., Baker E. N., Rumball S. V. Purification and preliminary crystallographic studies on azurin and cytochrome c' from Alcaligenes denitrificans and Alcaligenes sp. NCIB 11015. J Mol Biol. 1979 Nov 25;135(1):309–312. doi: 10.1016/0022-2836(79)90357-7. [DOI] [PubMed] [Google Scholar]
  14. Petratos K., Banner D. W., Beppu T., Wilson K. S., Tsernoglou D. The crystal structure of pseudoazurin from Alcaligenes faecalis S-6 determined at 2.9 A resolution. FEBS Lett. 1987 Jun 29;218(2):209–214. doi: 10.1016/0014-5793(87)81048-7. [DOI] [PubMed] [Google Scholar]
  15. Ross G. W., O'Callaghan C. H. Beta-lactamase assays. Methods Enzymol. 1975;43:69–85. doi: 10.1016/0076-6879(75)43081-6. [DOI] [PubMed] [Google Scholar]
  16. SAITO H., MIURA K. I. PREPARATION OF TRANSFORMING DEOXYRIBONUCLEIC ACID BY PHENOL TREATMENT. Biochim Biophys Acta. 1963 Aug 20;72:619–629. [PubMed] [Google Scholar]
  17. 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]
  18. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  19. Wallace R. B., Johnson M. J., Hirose T., Miyake T., Kawashima E. H., Itakura K. The use of synthetic oligonucleotides as hybridization probes. II. Hybridization of oligonucleotides of mixed sequence to rabbit beta-globin DNA. Nucleic Acids Res. 1981 Feb 25;9(4):879–894. doi: 10.1093/nar/9.4.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. de Boer H. A., Comstock L. J., Vasser M. The tac promoter: a functional hybrid derived from the trp and lac promoters. Proc Natl Acad Sci U S A. 1983 Jan;80(1):21–25. doi: 10.1073/pnas.80.1.21. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES