Specific overproduction and purification of the cytochrome b558 component of the cytochrome d complex from Escherichia coli

Green, George N.; Lorence, Robert M.; Gennis, Robert B.

doi:10.1021/bi00357a002

Cited by 56 publications

(47 citation statements)

References 10 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Cells containing pNG2, the pBR322-derived plasmid with the cloned cyd operon (17), are bright green because of the heme d component in the overproduced oxidase. Cells containing pNG10 are red since they only produce subunit I, which is the cytochrome b558 component of the oxidase (18). About 40% of the plasmids did not overexpress cytochrome and were not characterized further.…”

Section: Resultsmentioning

confidence: 99%

“…Briefly, cells were grown in 1.5 ml of LB-glucose plus 10 ,ug of tetracycline per ml for 12 to 24 h before harvest. After isolation, plasmids were resuspended in 20 ,ul of 10 mM Tris-HCl (Sigma)-1 mM EDTA (Sigma) (pH 8), and 15 ,ul was used to transform GR84N or G0102.…”

Section: Methodsmentioning

confidence: 99%

“…The antibody binding and proteolysis experiments also suggest that the site of ubiquinol oxidation is near the periplasmic side of the cytoplasmic membrane, consistent with a scalar mechanism of proton translocation (10,30). In addition, genetic studies have shown that the cytochrome b558 moiety is in subunit I (15). One of the axial ligands to cytochrome b558 has been identified by sitedirected mutagenesis to be His-186 in subunit I (11).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Isolation and characterization of a new class of cytochrome d terminal oxidase mutants of Escherichia coli

Oden

Gennis

1991

J Bacteriol

View full text Add to dashboard Cite

Cytochrome d terminal oxidase mutants were isolated by using hydroxylamine mutagenesis of pNG2, a pBR322-derived plasmid containing the wild-type cyd operon. The mutagenized plasmid was transformed into a cyo cyd recA strain, and the transformants were screened for the inability to confer aerobic growth on nonfermentable carbon sources. Western blot analysis and visible-light spectroscopy were performed to characterize three independent mutants grown both aerobically and anaerobically. The mutational variants of the cytochrome d complex were stabilized under anaerobic growth conditions. All three mutations perturb the b595 and d heme components of the complex. These mutations were mapped and sequenced and are shown to be located in the N-terminal third of subunit II of the cytochrome d complex. It is proposed that the N terminus of subunit II may interact with subunit I to form an interface that binds the b595 and d heme centers.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Isolation and characterization of a new class of cytochrome d terminal oxidase mutants of Escherichia coli

Oden

Gennis

1991

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…All three of the heme prosthetic groups are located on the periplasmic side of the membrane. Heme b 558 is known to be located entirely within subunit I (Green et al 1984). The Q-loop in subunit I has been suggested to participate in the binding of heme d and subunit II is necessary to bind heme b 595 and heme d in E. coli (Green et al 1984(Green et al , 1986.…”

Section: Introductionmentioning

confidence: 99%

Asymmetrical Evolution of Cytochrome bd Subunits

Hao

Golding

2006

J Mol Evol

View full text Add to dashboard Cite

Abstract. Functionally linked genes generally evolve at similar rates and the knowledge of this particular feature of genomic evolution has been used as the basis for the phylogenetic profiling method. We illustrate here an exception to this rule in the evolution of the cytochrome bd complex. This is a twocomponent oxidase complex, with the subunits I and II known to be widely present in bacteria. The subunits within the cytochrome bd complex are under the same evolutionary pressure and most likely behave in the same evolutionary manner. However, the sequence similarity of genes encoding subunit II varies considerably across species. Genes encoding subunit II evolve 1.2 times faster on most of the branches of their phylogeny than subunit I genes. Furthermore, the genes encoding subunit II in Oceanobacillus iheyensis, Bacillus halodurans, and Staphylococcus species do not have detectable homologues within E. coli due to their large divergence. Together, the two subunits of cytochrome bd reveal an interesting example of an asymmetric pattern of evolutionary change.

show abstract

“…The sequence identified showed 85% identity to that of the E. coli cydA gene (14), which codes for subunit I of the cytochrome d oxidase, and indicated that the TnphoA gene had inserted immediately after nucleotide 1478 of the putative S. typhimurium cydA ORF. E. coli cyd mutants show increased sensitivity to sodium azide and hydrogen peroxide, and they produce colonies that are the same size as those of the wild type when grown at 30°C but produce microcolonies at 37°C on L agar (15,31). Mutant GM129 showed an increased sensitivity to sodium azide, but not to hydrogen peroxide, and produced smaller colonies at 30, 37, and 42°C on L agar than those produced by the parent.…”

mentioning

confidence: 99%

Influence of genes encoding proton-translocating enzymes on suppression of Salmonella typhimurium growth and colonization

Zhang-Barber¹,

Turner²,

Martin³

et al. 1997

J Bacteriol

View full text Add to dashboard Cite

Twenty-four-hour-old, aerobically grown, Luria-Bertani broth cultures of Salmonella typhimurium F98 suppressed the growth of a spectinomycin-resistant (Spc r ) derivative of the same strain inoculated at 10 3 CFU ml ؊1 . This growth suppression is genus specific and RpoS independent, and it is not solely a result of nutrient depletion (P. A. Barrow, M. A. Lovell, and L. Zhang-Barber, J. Bacteriol. 178:3072-3076, 1996). Mutations in three genes are shown here to significantly reduce growth suppression under these conditions. The mutations were located in the nuo, cyd, and unc operons, which code for the NADH dehydrogenase I, cytochrome d oxidase, and F 0 F 1 proton-translocating ATPase complexes, respectively. When cultures were grown under strictly anaerobic conditions, only the unc mutant did not suppress growth. Prior colonization of the alimentary tract of newly hatched chickens with the S. typhimurium F98 wild type or nuo or cyd mutants suppressed colonization by an S. typhimurium F98 Spc r derivative inoculated 24 h later. In contrast, the S. typhimurium unc mutant did not suppress colonization. The nuo and unc mutants showed poorer growth on certain carbon sources. The data support the hypothesis that growth suppression operates because of the absence of a utilizable carbon source or electron acceptor.The normal microbial flora of the alimentary tract is a major factor protecting animals and humans against colonization by pathogenic bacteria (17). Newly hatched chickens without an established flora are particularly susceptible to Salmonella infection and colonization. Colonized young chicks often excrete large numbers of salmonellae into their environment for long periods, thus becoming a potential reservoir of infection (7). When chickens reach several weeks of age, the lower alimentary tract is colonized by large numbers of obligate anaerobes which confer increased resistance to the establishment of Salmonella infection. The resistance of young chicks to infection may be artificially increased by oral administration, immediately after hatching, of a suspension or culture of cecal contents obtained from healthy adult birds (23). This phenomenon has been termed "competitive exclusion," and its discovery has led to a search for individual strains of bacteria which can inhibit colonization by pathogens to the same extent as the normal flora. Oral administration of an avirulent Salmonella typhimurium strain to day-old chicks can confer protection against oral infection by a virulent strain (known as colonization inhibition) (8, 10). Colonization inhibition is normally genus specific, is not caused by bacteriophage or bacteriocin activity, and is not a consequence of immunity induced by the first strain (8). Attempts are currently under way to elucidate the mechanisms responsible for this effect. An analogous phenomenon in vitro has been observed with 24-h-old stationaryphase nutrient broth cultures of S. typhimurium (6,8,11). When such cultures are inoculated with small numbers of genetically tagged (antibiot...

show abstract

Specific overproduction and purification of the cytochrome b558 component of the cytochrome d complex from Escherichia coli

Cited by 56 publications

References 10 publications

Isolation and characterization of a new class of cytochrome d terminal oxidase mutants of Escherichia coli

Isolation and characterization of a new class of cytochrome d terminal oxidase mutants of Escherichia coli

Asymmetrical Evolution of Cytochrome bd Subunits

Influence of genes encoding proton-translocating enzymes on suppression of Salmonella typhimurium growth and colonization

Contact Info

Product

Resources

About