Topological studies of monomeric and dimeric cytochrome c oxidase and identification of the copper A site using a fluorescence probe.

Hall, Joan; Moubarak, Ali S.; O'Brien, P; Pan, Lian Ping; Cho, I.; Millett, Francis

doi:10.1016/s0021-9258(18)68453-4

Cited by 31 publications

(9 citation statements)

References 43 publications

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“…Hall et al have studied the topology of cytochrome oxidase by Forster energy transfer between an extrinsic fluorescence probe bound to subunit III and the heme of cytochrome c bound to the high affinity site on cytochrome oxidase. They concluded that in cytochrome oxidase dimers, subunit III of one monomer is relatively close (26-40 A) to the cytochrome c binding site on subunit II of the other monomer, but within one monomer subunit III is more than 54 A from the cytochrome c binding site (Hall et al, 1988). This supports the model for cytochrome c binding to cytochrome oxidase dimers proposed by Capaldi et al…”

Section: Discussionsupporting

confidence: 61%

“…Cys-115 of subunit III is the sulfhydryl in cytochrome oxidase most reactive to hydrophilic sulfhydryl reagents, and Cys-115 can be labeled selectively under appropriate conditions (McGeer et al, 1977;Hall et al, 1988;Malatesta & Capaldi, 1982; Muller & Azzi, 1985). Yang et al (1984) have described the synthesis of an undecagold cluster compound containing a single maleimide group which can be used to label selectively cysteine sulfhydryl groups.…”

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confidence: 99%

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Electron Microscopy of Cytochrome c Oxidase Crystals: Labeling of Subunit III with a Monomaleimide Undecagold Cluster Compound

Crum

Gruys²,

Frey³

1994

Biochemistry

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Two-dimensional crystals of beef heart mitochondrial cytochrome c oxidase dimers were labeled at Cys-115 of subunit III with a monomaleimide derivative of an undecagold cluster compound. The binding site of the gold cluster compound and hence the site of subunit III were identified by image processing of cryoelectron micrographs of the crystals preserved in a mixture of glucose and uranyl acetate. The shape of the cytochrome oxidase dimer can be approximated as a parallelogram which is 44 by 82 A with an included angle of 80 degrees oriented with its long dimension along the a axis of the crystal. Labeling of subunit III was confirmed by a shift in the mobility of approximately 50% of subunit III molecules upon electrophoresis in polyacrylamide gels in the presence of sodium dodecyl sulfate. Averaged images of undecagold cluster labeled crystals and of unlabeled crystals were calculated; each image represents an average of approximately 17,000 molecules of either labeled or unlabeled cytochrome oxidase. On the basis of a statistical analysis of the differences between the two images, the gold cluster binds along a line 30 degrees from the a axis and 29 A from the center of the dimer. This result is interpreted in the context of other structural studies including the site of cytochrome c binding which Frey and Murray found to be near the a axis and 18 A from the center of the dimer [Frey, T. G., & Murray, J. M. (1994) J. Mol. Biol. 237, 275-297].(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Discussionsupporting

confidence: 61%

mentioning

confidence: 99%

Electron Microscopy of Cytochrome c Oxidase Crystals: Labeling of Subunit III with a Monomaleimide Undecagold Cluster Compound

Crum

Gruys²,

Frey³

1994

Biochemistry

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“…No inhibition is observed if the cytochrome 0 complex is incorporated into proteoliposomes. The inhibition data, however, demonstrate that although the role of subunit II in the quinol oxidase must be different from its presumed role in the cytochrome c oxidases, i.e., interacting with cytochrome c and binding to CuA (Bisson et al, 1982;Capaldi et al, 1987;Covello & Gray, 1990;Hall et al, 1988;Holm et al, 1987), it still must play some critical function in the cytochrome o complex.…”

Section: Resultsmentioning

confidence: 81%

“…In addition to the copper within the binuclear center (Cub), the cytochrome c oxidases contain at least one other copper (CuA), which is EPRdetectable and is involved in the oxidation of cytochrome c (Chan & Li, 1990;Li et al, 1987;Morgan etal., 1989;Stevens et al, 1982). Sequence comparisons and biochemical studies suggest that this copper (CuA) is located within subunit II of the cytochrome c oxidases (Capaldi et al, 1987;Covello & Gray, 1990;Hall et al, 1988;Holm et al, 1987;Li et al, 1987;Saraste, 1990). It has been proposed that this EPR signal is due to a two-copper center (Kroneck et al, 1990), although this has been disputed (Pan etal., 1991).…”

mentioning

confidence: 99%

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

Kc¹,

Vc²,

Ne³

et al. 1992

Biochemistry

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The cytochrome o complex is a bo-type ubiquinol oxidase in the aerobic respiratory chain of Escherichia coli. This complex has a close structural and functional relationship with the eukaryotic and prokaryotic aa3-type cytochrome c oxidases. The specific activity, subunit composition, and metal content of the purified cytochrome o complex are not consistent for different preparative protocols reported in the literature. This paper presents a relatively simple preparation of the enzyme starting with a strain of Escherichia coli which overproduces the oxidase. The pure enzyme contains four subunits by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Partial amino acid sequence data confirm the identities of subunit I, II, and III from the SDS-PAGE analysis as the cyoB, cyoA, and cyoC gene products, respectively. A slight modification of the purification protocol yields an oxidase preparation that contains a possible fifth subunit which may be the cyoE gene product. The pure four-subunit enzyme contains 2 equivs of iron but only 1 equiv of copper. There is no electron paramagnetic resonance detectable copper in the purified enzyme. Hence, the equivalent of CuA of the aa3-type cytochrome c oxidases is absent in this quinol oxidase. There is also no zinc in the purified quinol oxidase. Finally, monoclonal antibodies are reported that interact with subunit II. One of these monoclonals inhibits the quinol oxidase activity of the detergent-solubilized, purified oxidase. Hence, although subunit II does not contain CuA and does not interact with cytochrome c, it still must have an important function in the bo-type ubiquinol oxidase.

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“…Accordingly, we have proposed that CuA is ligated by two cysteine sulfur atoms and two histidine nitrogen atoms. Comparison of amino acid sequences for cytochrome oxidases across a wide variety of organisms does show the presence of two highly conserved cysteine residues in subunit II (Steffens et al, 1987;Hall et al, 1988), and it has been surmised that these are the two cysteine ligands to CuA. No other cysteine residues are conserved.…”

Section: Structural Biochemistrymentioning

confidence: 99%

Cytochrome c oxidase: understanding nature's design of a proton pump

Chan¹,

Li²

1990

Biochemistry

194

117

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Topological studies of monomeric and dimeric cytochrome c oxidase and identification of the copper A site using a fluorescence probe.

Cited by 31 publications

References 43 publications

Electron Microscopy of Cytochrome c Oxidase Crystals: Labeling of Subunit III with a Monomaleimide Undecagold Cluster Compound

Electron Microscopy of Cytochrome c Oxidase Crystals: Labeling of Subunit III with a Monomaleimide Undecagold Cluster Compound

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

Cytochrome c oxidase: understanding nature's design of a proton pump

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