Modified, large-scale purification of the cytochrome o complex (bo-type oxidase) of Escherichia coli yields a two heme/one copper terminal oxidase with high specific activity

Kc, Minghetti; Vc, Goswitz; Ne, Gabriel; Jj, Hill; Ca, Barassi; Georgiou, Christos D.; Si, Chan; Rb, Gennis

doi:10.1021/bi00145a008

Cited by 75 publications

(59 citation statements)

References 75 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…We feel that such a possibility exists and that there are several reasons why additional eubacterial accessory polypeptides have not been discovered. One of the foremost arguments against the existence of eubacterial accessory factors is the observation that biochemical purification of the eubacterial enzyme complex does not show the presence of the numerous accessory polypeptides that are observed in similar enzyme preparations obtained from yeast and mammalian mitochondria (27,29,33,46). This discrepancy, however, can be rationalized by assuming either that eubacterial accessory polypeptides are not associated with the mature holoenzyme complex or that they are loosely associated and subsequently stripped off during detergent-mediated solubilization and purification of the enzyme complex.…”

Section: Discussionmentioning

confidence: 99%

Cloning and characterization of senC, a gene involved in both aerobic respiration and photosynthesis gene expression in Rhodobacter capsulatus

Buggy

Bauer

1995

J Bacteriol

View full text Add to dashboard Cite

The purple nonsulfur photosynthetic eubacterium Rhodobacter capsulatus is a versatile organism that can obtain cellular energy by several means, including the capture of light energy for photosynthesis as well as the use of light-independent respiration, in which molecular oxygen serves as a terminal electron acceptor. In this study, we have identified and characterized a novel gene, senC, mutations in which affect respiration as well as the induction of photosynthesis gene expression. The protein coded by senC exhibits 33% sequence identity to the yeast nucleus-encoded protein SCO1, which is thought to be a mitochondrion-associated cytochrome c oxidase assembly factor. Like yeast SCO1, SenC is required for optimal cytochrome c oxidase activity in aerobically grown R. capsulatus cells. We further show that senC is required for maximal induction from the puf and puh operons, which encode the structural polypeptides of the light-harvesting and reaction center complexes.

show abstract

Section: Discussionmentioning

confidence: 99%

Cloning and characterization of senC, a gene involved in both aerobic respiration and photosynthesis gene expression in Rhodobacter capsulatus

Buggy

Bauer

1995

J Bacteriol

View full text Add to dashboard Cite

show abstract

“…The WT Pp and morA Pp mutant strains were transformed with pGB3 (5) expressing green fluorescent protein (GFP). Crude membrane preparations were prepared from both WT Pp and the morA Pp mutant by using a previously described method (12). Anti-MorA polyclonal antibodies against part of MorA Pp (amplified with primers MorPASB [5Ј-CCT CCC GGA TCC CTG ATC GAG CTG TCG TTG C-3Ј] and MorENDH [5Ј-CCT CCC AAG CTT TTA GTC GAA CAT GAA CAG CGC-3Ј]) were raised in rabbits.…”

Section: Methodsmentioning

confidence: 99%

MorA Defines a New Class of Regulators Affecting Flagellar Development and Biofilm Formation in Diverse Pseudomonas Species

Choy¹,

et al. 2004

View full text Add to dashboard Cite

Assembly of bacterial flagella is developmentally important during both planktonic cell growth and biofilm formation. Flagellar biogenesis is complex, requiring coordinated expression of over 40 genes, and normally commences during the log-to-stationary transition phase. We describe here a novel membrane-localized regulator, MorA, that controls the timing of flagellar development and affects motility, chemotaxis, and biofilm formation in Pseudomonas putida. MorA is conserved among diverse Pseudomonas species, and homologues are present in all Pseudomonas genomes sequenced thus far. In P. putida, the absence of MorA derepresses flagellar development, which leads to constitutive formation of flagella in the mutant cells in all growth phases. In Pseudomonas aeruginosa, the absence of MorA led to a reduction in biofilm formation. However, unlike the motility of P. putida, the motility of the P. aeruginosa mutants was unaffected. Our data illustrate a novel developmentally regulated sensory and signaling pathway for several properties required for virulence and ecological fitness of Pseudomonas species.Bacterial cells exist either as free-swimming planktonic cells or in surface-attached communities known as biofilms. In planktonic cell cultures, the log-to-stationary transition phase is marked by the onset of flagellar development, which is a highly complex process involving coordinated expression of over 40 genes in a hierarchical manner (3,8,11,16). Recently, the regulatory control of flagellar biogenesis in Pseudomonas aeruginosa was described in detail (7). In contrast to the threetier regulation in the multiflagellated organism Escherichia coli, a four-tier hierarchy is present in the monoflagellated organism P. aeruginosa. At the top of the hierarchy are the transcriptional regulators, FleQ, and the alternative sigma factor FliA, which regulate at least 11 operons. Planktonic cells undergo multiple developmental changes during their transition from free-swimming organisms to the surface-attached bacterial communities that constitute biofilms. Appropriate levels of flagellin subunits seem to be a key factor since overexpression of flagellin in E. coli results in reduced adhesion (10). In fully developed biofilms, bacteria like Pseudomonas putida may even lack flagella (22). However, in P. aeruginosa, the role of flagella in biofilm formation still remains an open question as a recent study of biofilm development showed that flagella are not involved in the attachment and that initial microcolony formation occurs by clonal growth (9). Despite the fact that there is a great deal of knowledge concerning the flagellar pathway in diverse bacteria, very little is known about the negative regulation of this process.Here we describe identification of MorA, a membrane-localized negative regulator of the timing of flagellar formation. In the mutilflagellated organism P. putida, planktonic cells of a morA mutant have constitutive expression of flagella. This property enhances motility and chemotaxis, but it interferes with...

show abstract

“…In contrast, due to the extreme hydrophobic nature of the physiological substrate ubiquinol-8, the ubiquinol binding site(s) is (are) expected to exist in the transmembrane region of the ubiquinol oxidase complexes as is analogously observed in the structures of bacterial photosynthetic reaction centers. If a ubiquinol binding domain exists on subunit II of the cytochrome bo, complex, as has been suggested by a number of studies [4,5], the substrate oxidizing domains of the two families of terminal oxidases would exist in different three-dimensional locations and protein environments of subunit II. Thus, the subunit II's of the two families of oxidases could have either very different folding patterns to accommodate the different substrates or they could have very similar overall folding patterns for evolutionary reasons.…”

mentioning

confidence: 95%

Further comparison of ubiquinol and cytochrome c terminal oxidases

1993

Self Cite

View full text Add to dashboard Cite

In the preceeding commentary, Haltia reflects on our view presented in [l] that the ubiquinol and cytochrome c terminal oxidases are likely to utilize different electron transfer and proton translocation mechanisms. We would like to take this opportunity to clarify our position on some of the issues that he discusses.Haltia compares the structure and function of subunit II of the two families of terminal oxidases, the ubiquinol oxidases and the cytochrome c oxidases. While structure prediction in general is hardly a mature field, hydrophobicity calculations are relatively accurate for predicting the number and approximate location of transmembrane helices in a membrane protein. However, such predictions are inaccurate at times [2,3] and, moreover, say virtually nothing about the extramembraneous domains. To predict that two polypeptides have a similar structure on the basis of hydrophobicity calculations is pushing the technique beyond the limits of its capabilities. As pointed out in our minireview [l], the Cu, site and part or all of the residues responsible for the binding of cytochrome c are thought to be located in the extra-membraneous domain of subunit II of the cytochrome c oxidases. In contrast, due to the extreme hydrophobic nature of the physiological substrate ubiquinol-8, the ubiquinol binding site(s) is (are) expected to exist in the transmembrane region of the ubiquinol oxidase complexes as is analogously observed in the structures of bacterial photosynthetic reaction centers. If a ubiquinol binding domain exists on subunit II of the cytochrome bo, complex, as has been suggested by a number of studies [4,5], the substrate oxidizing domains of the two families of terminal oxidases would exist in different three-dimensional locations and protein environments of subunit II. Thus, the subunit II's of the two families of oxidases could have either very different folding patterns to accommodate the different substrates or they could have very similar overall folding patterns for evolutionary reasons. We note that the 10% sequence identity of subunit II of the Escherichia coli cytochrome bo, complex and the Paracoccus denitrz@ans cytochrome au, complex is well below what Doolittle terms the 'twilight zone' (15-25% sequence identity), and accordingly, these polypeptides are unlikely to be evolutionarily related based on sequence comparison alone [6]. Abbreviations: HQNO, 2-heptyl4hydroxyquinoline N-oxide, UHDBT, Haltia supports his argument for similar overall folding patterns for subunit II of the two families of oxidases with experiments in which up to six residues in a fragment of the E. coli quinol oxidase subunit II were mutated in an attempt to create a Cu, site [7,8]. It remains to be shown that the peptide conformations with and without the mutated residues are basically the same. Virtually any polypeptide into which cysteines and histidines are introduced will bind copper, most likely in an altered conformation because of the nucleating effect of the metal ions. Van der Oost and coworkers found ...

show abstract

Modified, large-scale purification of the cytochrome o complex (bo-type oxidase) of Escherichia coli yields a two heme/one copper terminal oxidase with high specific activity

Cited by 75 publications

References 75 publications

Cloning and characterization of senC, a gene involved in both aerobic respiration and photosynthesis gene expression in Rhodobacter capsulatus

Cloning and characterization of senC, a gene involved in both aerobic respiration and photosynthesis gene expression in Rhodobacter capsulatus

MorA Defines a New Class of Regulators Affecting Flagellar Development and Biofilm Formation in Diverse Pseudomonas Species

Further comparison of ubiquinol and cytochrome c terminal oxidases

Contact Info

Product

Resources

About