Characterization of the Cytochrome <i>c</i> Membrane‐Binding Site Using Cardiolipin‐Containing Bicelles with NMR

Kobayashi, Hisashi; Nagao, Shigeaki; Hirota, Shun

doi:10.1002/ange.201607419

Cited by 5 publications

(3 citation statements)

References 40 publications

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“…[4a] Notwithstanding the different crystallization conditions,the larger pclx 6 (ammonium sulfate precipitant) was more specific than sclx 4 (PEG precipitant). [23] Finally, we note that the pclx 6 -cyt c complex provides structural clues on the interactions of cyt c with phospholipids in the mitochrondrial membrane.…”

Section: Angewandte Chemiementioning

confidence: 99%

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Protein Dimerization on a Phosphonated Calix[6]arene Disc

Rennie,

Doolan,

Raston

et al. 2017

Angew Chem Int Ed

110

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Complex formation between cationic cytochrome c and the water-soluble, poly-anionic p-phosphonatocalix[6]arene (pclx ) was investigated. A crystal structure (at 1.8 Å resolution) revealed a remarkable dimeric disc of pclx that acts like glue to mediate a symmetric (C ) protein dimer. The calixarene disc has a diameter of about 1.5 nm and masks about 360 Å of protein surface. The key protein-calixarene contacts occur via two linchpin lysines, with additional contacts provided by a small hydrophobic patch. The protein-calixarene supramolecular assemblies were observed in solution by size-exclusion chromatography with multi-angle light scattering and NMR spectroscopy. Using isothermal titration calorimetry and NMR data, an apparent K in the low micromolar range was determined for the charge-rich protein-calixarene complex. In contrast to p-sulfonatocalix[4]arene, the larger pclx has a single, well-defined binding site that mediates the assembly of cytochrome c in solution.

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Section: Angewandte Chemiementioning

confidence: 99%

“…Finally, we note that the pclx 6 -cyt c complex provides structural clues on the interactions of cyt c with phospholipids in the mitochrondrial membrane. [23]…”

Section: [4a-c]mentioning

confidence: 99%

Protein Dimerization on a Phosphonated Calix[6]arene Disc

Rennie,

Doolan,

Raston

et al. 2017

Angew Chem Int Ed

110

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show abstract

“…NMR spectroscopy can simultaneously identify and monitor different molecules or conformers "without prejudice" and provide protein structure information at atomic resolution. It has been successfully used to detect the binding site of cyt c upon interaction with CL [21,22]. However, it is difficult to be used to observe structural changes at a high CL/cyt c molar ratio, as cyt c, when bound to large, slow-tumbling liposomes, has a higher correlation time, causing a decrease in NMR signal intensity [23,24], which, in turn, results in the disappearance of 1 H- 15 N HSQC signals of cyt c upon interaction with high content of CL [22].…”

Section: Introductionmentioning

confidence: 99%

NMR Reveals the Conformational Changes of Cytochrome C upon Interaction with Cardiolipin

Zhan

Zhang

Chai

et al. 2021

Life

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Conformational change of cytochrome c (cyt c) caused by interaction with cardiolipin (CL) is an important step during apoptosis, but the underlying mechanism is controversial. To comprehensively clarify the structural transformations of cyt c upon interaction with CL and avoid the unpredictable alias that might come from protein labeling or mutations, the conformation of purified yeast iso–1 cyt c with natural isotopic abundance in different contents of CL was measured by using NMR spectroscopy, in which the trimethylated group of the protein was used as a natural probe. The data demonstrate that cyt c has two partially unfolded conformations when interacted with CL: one with Fe–His33 coordination and the other with a penta–coordination heme. The Fe–His33 coordination conformation can be converted into a penta–coordination heme conformation in high content of CL. The structure of cyt c becomes partially unfolded with more exposed heme upon interaction with CL, suggesting that cyt c prefers a high peroxidase activity state in the mitochondria, which, in turn, makes CL easy to be oxidized, and causes the release of cyt c into the cytoplasm as a trigger in apoptosis.

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Protein Dimerization on a Phosphonated Calix[6]arene Disc

et al. 2017

View full text Add to dashboard Cite

Complex formation between cationic cytochrome c and the water‐soluble, poly‐anionic p‐phosphonatocalix[6]arene (pclx6) was investigated. A crystal structure (at 1.8 Å resolution) revealed a remarkable dimeric disc of pclx6 that acts like glue to mediate a symmetric (C2) protein dimer. The calixarene disc has a diameter of about 1.5 nm and masks about 360 Å2 of protein surface. The key protein–calixarene contacts occur via two linchpin lysines, with additional contacts provided by a small hydrophobic patch. The protein–calixarene supramolecular assemblies were observed in solution by size‐exclusion chromatography with multi‐angle light scattering and NMR spectroscopy. Using isothermal titration calorimetry and NMR data, an apparent Kd in the low micromolar range was determined for the charge‐rich protein–calixarene complex. In contrast to p‐sulfonatocalix[4]arene, the larger pclx6 has a single, well‐defined binding site that mediates the assembly of cytochrome c in solution.

show abstract

Characterization of the Cytochrome c Membrane‐Binding Site Using Cardiolipin‐Containing Bicelles with NMR

Cited by 5 publications

References 40 publications

Protein Dimerization on a Phosphonated Calix[6]arene Disc

Protein Dimerization on a Phosphonated Calix[6]arene Disc

NMR Reveals the Conformational Changes of Cytochrome C upon Interaction with Cardiolipin

Protein Dimerization on a Phosphonated Calix[6]arene Disc

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