Investigation of the essential boundary layer phospholipids of cytochrome c oxidase using Triton X-100 delipidation

Robinson, N.; Strey, Fredlein; Talbert, Linda

doi:10.1021/bi00557a003

Cited by 199 publications

(112 citation statements)

References 22 publications

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“…Interestingly, the two high-affinity CL binding sites were located in close proximity to the proton channels and were associated with the regulation of the enzyme (32,33), whereas the low-affinity binding sites were linked to stabilization of subunits VIa and VIb, important for dimerization of CcO (30,34). A loss of CcO function in the absence of CL was shown recently through destabilization of quaternary structure following CL removal (13,14).…”

Section: Discussionmentioning

confidence: 97%

Dimer interface of bovine cytochrome c oxidase is influenced by local posttranslational modifications and lipid binding

Liko

Degiacomi

Mohammed

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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Bovine cytochrome c oxidase is an integral membrane protein complex comprising 13 protein subunits and associated lipids. Dimerization of the complex has been proposed; however, definitive evidence for the dimer is lacking. We used advanced mass spectrometry methods to investigate the oligomeric state of cytochrome c oxidase and the potential role of lipids and posttranslational modifications in its subunit interfaces. Mass spectrometry of the intact protein complex revealed that both the monomer and the dimer are stabilized by large lipid entities. We identified these lipid species from the purified protein complex, thus implying that they interact specifically with the enzyme. We further identified phosphorylation and acetylation sites of cytochrome c oxidase, located in the peripheral subunits and in the dimer interface, respectively. Comparing our phosphorylation and acetylation sites with those found in previous studies of bovine, mouse, rat, and human cytochrome c oxidase, we found that whereas some acetylation sites within the dimer interface are conserved, suggesting a role for regulation and stabilization of the dimer, phosphorylation sites were less conserved and more transient. Our results therefore provide insights into the locations and interactions of lipids with acetylated residues within the dimer interface of this enzyme, and thereby contribute to a better understanding of its structure in the natural membrane. Moreover dimeric cytochrome c oxidase, comprising 20 transmembrane, six extramembrane subunits, and associated lipids, represents the largest integral membrane protein complex that has been transferred via electrospray intact into the gas phase of a mass spectrometer, representing a significant technological advance.cytochrome c oxidase | mass spectrometry | lipids

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Section: Discussionmentioning

confidence: 97%

Dimer interface of bovine cytochrome c oxidase is influenced by local posttranslational modifications and lipid binding

Liko

Degiacomi

Mohammed

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

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“…A functional role for two or three molecules of (Ptd)2Gro in cytochrome c oxidase activity has been described recently (41).…”

Section: Discussionmentioning

confidence: 99%

Diphosphatidylglycerol is required for optimal activity of beef heart cytochrome c oxidase.

Vik¹,

Georgevich²,

Capaldi³

1981

Proc. Natl. Acad. Sci. U.S.A.

160

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Isolated beef heart cytochrome c oxidase (ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1) contains four or five molecules of tightly bound diphosphatidylglycerol per monomer (2-heme complex). This lipid could be removed in part, or wholly, by mixing the enzyme with high concentrations of Triton X-100 and then centrifuging the mixture through a glycerol gradient equilibrated in the same detergent. Cytochrome c oxidase retaining three or more diphosphatidylglycerol molecules per monomer was fully active when assayed in 1-oleoyl lysophosphatidylcholine. Upon removal of one or more of these diphosphatidylglycerols, enzymic activity was lost. Full activation could be obtained by adding diphosphatidylglycerol to the assay mixture along with lysophosphatidylcholine but not by adding phosphatidylcholine or phosphatidylethanolamine. Direct binding experiments, kinetic studies, and previous work using arylazidocytochrome c derivatives [Bisson, R., Jacobs, B. & Capaldi, R. A. (1980) Biochemistry 10, 4173-4178], indicate that diphosphatidylglycerol is involved in binding of substrate cytochrome c to cytochrome c oxidase.

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“…As these cardiolipin molecules are progressively removed, the number of cytochrome c molecules bound is reduced to one and the low affinity phase of electron transfer also disappears ( fig.2). These results indicate that negatively charged phospholipids contribute an Important part of the low affinity binding site (see also [46]). …”

Section: Structure Of Cytochrome C Binandg Sites On Cytochrome C Oxidasementioning

confidence: 65%

Structural and functional features of the interaction of cytochrome c with complex III and cytochrome c oxidase

et al. 1982

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Investigation of the essential boundary layer phospholipids of cytochrome c oxidase using Triton X-100 delipidation

Cited by 199 publications

References 22 publications

Dimer interface of bovine cytochrome c oxidase is influenced by local posttranslational modifications and lipid binding

Dimer interface of bovine cytochrome c oxidase is influenced by local posttranslational modifications and lipid binding

Diphosphatidylglycerol is required for optimal activity of beef heart cytochrome c oxidase.

Structural and functional features of the interaction of cytochrome c with complex III and cytochrome c oxidase

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