Home LiteratureArticle Details
PMID: 1655703 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Cloning, characterization, and expression in Escherichia coli of the genes encoding the cytochrome d oxidase complex from Azotobacter vinelandii.

Journal of bacteriology ·Vol. 173 ·No. 19 ·1991-10-00 ·Pages 6230-41

Moshiri F, Chawla A, Maier RJ

Abstract

Azotobacter vinelandii is a free-living nitrogen-fixing bacterium that has one of the highest respiratory rates of all aerobic organisms. Based on various physiological studies, a d-type cytochrome has been postulated to be the terminal oxidase of a vigorously respiring but apparently uncoupled branch of the electron transport system in the membranes of this organism. We cloned and characterized the structural genes of the two subunits of this oxidase. The deduced amino acid sequences of both subunits of the A. vinelandii oxidase have extensive regions of homology with those of the two subunits of the Escherichia coli cytochrome d complex. Most notably, the histidine residues proposed to be the axial ligands for the b hemes of the E. coli oxidase and an 11-amino-acid stretch proposed to be part of the ubiquinone binding site are all conserved in subunit I of the A. vinelandii oxidase. The A. vinelandii cytochrome d was expressed in a spectrally and functionally active form in the membranes of E. coli, under the control of the lac or tac promoter. The spectral features of the A. vinelandii cytochrome d expressed in E. coli are very similar to those of the E. coli cytochrome d. The expressed oxidase was active as a quinol oxidase and could reconstitute an NADH to oxygen electron transport chain.

Related Genes
cyd
MeSH Terms
Amino Acid Sequence Azotobacter vinelandii/enzymology,genetics Base Sequence Cloning, Molecular Cytochrome b Group Cytochrome d Group Cytochromes/genetics Electron Transport Chain Complex Proteins Escherichia coli/enzymology,genetics Escherichia coli Proteins Genes, Bacterial Genetic Vectors Molecular Sequence Data Oxidoreductases/biosynthesis,genetics Sequence Homology, Nucleic Acid
Chemicals
Cytochrome b Group Cytochromes Electron Transport Chain Complex Proteins Escherichia coli Proteins Cytochrome d Group Oxidoreductases cytochrome bd terminal oxidase complex, E coli
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Moshiri F
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
Chawla A
Maier R J
References (29)
29 references, click to expand
  1. The 3'-terminal sequence of Escherichia coli 16S ribosomal RNA: complementarity to nonsense triplets and ribosome binding sites.
    Proc Natl Acad Sci U S A. 1974 Apr;71(4):1342-6 PMID: 4598299
  2. Electron transport in Azotobacter vinelandii.
    Biochim Biophys Acta. 1966 Mar 7;113(3):467-81 PMID: 4288128
  3. Preparation of red and green electron transport particles from Azotobacter vinelandii.
    Biochim Biophys Acta. 1967 Sep 6;143(2):354-62 PMID: 6049954
  4. Adaptation of Azotobacter cells to tricarboxylic acid substrates.
    J Bacteriol. 1954 Mar;67(3):353-60 PMID: 13143004
  5. Cloning and mutagenesis of genes encoding the cytochrome bd terminal oxidase complex in Azotobacter vinelandii: mutants deficient in the cytochrome d complex are unable to fix nitrogen in air.
    J Bacteriol. 1990 Oct;172(10):6010-9 PMID: 2170336
  6. Regulation of expression of the cytochrome d terminal oxidase in Escherichia coli is transcriptional.
    J Bacteriol. 1988 Feb;170(2):961-6 PMID: 2828338
  7. The nucleotide sequence of the cyd locus encoding the two subunits of the cytochrome d terminal oxidase complex of Escherichia coli.
    J Biol Chem. 1988 Sep 15;263(26):13138-43 PMID: 2843510
  8. Coulometric and spectroscopic analysis of the purified cytochrome d complex of Escherichia coli: evidence for the identification of "cytochrome a1" as cytochrome b595.
    Biochemistry. 1986 May 6;25(9):2314-21 PMID: 3013299
  9. "ATG vectors' for regulated high-level expression of cloned genes in Escherichia coli.
    Gene. 1985;40(2-3):183-90 PMID: 3007288
  10. Proposal that the function of the membrane-bound cytochrome a1-like haemoprotein (cytochrome b-595) in Escherichia coli is a direct electron donation to cytochrome d.
    FEBS Lett. 1987 Jun 8;217(1):49-52 PMID: 3036575
  11. Immunological investigation of the distribution of cytochromes related to the two terminal oxidases of Escherichia coli in other gram-negative bacteria.
    J Bacteriol. 1985 Feb;161(2):709-13 PMID: 2981822
  12. Bacterial electron transport chains.
    Annu Rev Biochem. 1988;57:101-32 PMID: 3052268
  13. Prediction and comparison of the haem-binding sites in membrane haemoproteins.
    Biochim Biophys Acta. 1989 Dec 7;977(3):249-65 PMID: 2686753
  14. Location of heme axial ligands in the cytochrome d terminal oxidase complex of Escherichia coli determined by site-directed mutagenesis.
    J Biol Chem. 1989 May 15;264(14):8026-32 PMID: 2656671
  15. Cytochrome o type oxidase from Escherichia coli. Characterization of the enzyme and mechanism of electrochemical proton gradient generation.
    Biochemistry. 1984 Sep 25;23(20):4703-14 PMID: 6093862
  16. Cloning the cyd gene locus coding for the cytochrome d complex of Escherichia coli.
    Gene. 1984 Dec;32(1-2):99-106 PMID: 6099317
  17. Epitopes of monoclonal antibodies which inhibit ubiquinol oxidase activity of Escherichia coli cytochrome d complex localize functional domain.
    J Biol Chem. 1990 Mar 15;265(8):4273-7 PMID: 1689724
  18. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
    Anal Biochem. 1983 Jul 1;132(1):6-13 PMID: 6312838
  19. Isolation and characterization of an Escherichia coli mutant lacking cytochrome d terminal oxidase.
    J Bacteriol. 1983 Jun;154(3):1269-75 PMID: 6304009
  20. The purification and characterization of the cytochrome d terminal oxidase complex of the Escherichia coli aerobic respiratory chain.
    J Biol Chem. 1983 Aug 10;258(15):9159-65 PMID: 6307994
  21. Hydrogen-oxidizing electron transport components in nitrogen-fixing Azotobacter vinelandii.
    J Bacteriol. 1984 Jul;159(1):348-52 PMID: 6735984
  22. Oxygen and hydrogen in biological nitrogen fixation.
    Annu Rev Microbiol. 1980;34:183-207 PMID: 6776883
  23. Evidence for an alternative nitrogen fixation system in Azotobacter vinelandii.
    Proc Natl Acad Sci U S A. 1980 Dec;77(12):7342-6 PMID: 6938981
  24. A simple method for displaying the hydropathic character of a protein.
    J Mol Biol. 1982 May 5;157(1):105-32 PMID: 7108955
  25. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.
    Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350-4 PMID: 388439
  26. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system.
    Gene. 1977;2(2):95-113 PMID: 344137
  27. The inhibition of Azotobacter vinelandii terminal oxidases by cyanide.
    FEBS Lett. 1973 Nov 1;36(3):347-50 PMID: 4357815
  28. The cytochrome system of Azotobacter vinelandii.
    Biochim Biophys Acta. 1967 Sep 6;143(2):340-53 PMID: 4292889
  29. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.173.19.6230-6241.1991
Published
1991-10-00
Pages
6230-41
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC208375
Subset
IM
Grants
NIGMS NIH HHS · GM40079 · United States
Databases
GENBANK
M59294, M71236, M71237, M71238, M71239, M71240, M71241, M88719, S56966, S57066
External Links
PubMed source DOI: 10.1128/jb.173.19.6230-6241.1991 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2