Binding of S. cerevisiae iso-1 cytochrome c and its surface lysine-to-alanine variants to cardiolipin: charge effects and the role of the lipid to protein ratio

Paradisi, Alessandro; Bellei, Marzia; Paltrinieri, Licia; Bortolotti, Carlo Augusto; Rocco, Giulia Di; Ranieri, Antonio; Borsari, Marco; Sola, Marcó; Battistuzzi, Gianantonio

doi:10.1007/s00775-020-01776-1

Cited by 13 publications

(22 citation statements)

References 104 publications

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“…As we have also found previously that the unfolding behavior of immobilized ycc changes with the nature of the adsorbing construct, 76,77 it is apparent how, from a more general perspective, the motional properties of the protein and the nature of the adsorbing surface constitute additional means that Nature can exploit to modulate protein unfolding and the eventual molecular response. 5,13,31…”

Section: Discussionmentioning

confidence: 99%

“…2c and d) indicates that a minor high-spin (HS1) form is also present. 13 For both variants, the Soret band in the absorption spectra and the single trough in the corresponding 2nd derivative spectra slightly redshift upon urea addition ( Fig. 2a-d and Table 1), whereas the shoulder in the 2nd derivative spectra at 398 nm disappears above 4 M urea ( Fig.…”

Section: Absorption and MCD Spectramentioning

confidence: 93%

“…The former were converted to molar ellipticity [y] using the conversion factor [y] = y(deg)Á100/(dÁc), where c is the protein concentration (mol dm À3 ) and d is the thickness of the sample (path length, 0.5 cm), 21,84,90 while the latter were converted to De [M À1 cm À1 T À1 ] using the conversion factor De = y/(32 980ÁcÁdÁB), where c is the protein concentration, B is the magnetic field (1 T), and d is the thickness of the sample (path length, 0.5 cm). 13,[101][102][103][104] All experiments were carried out at 25 1C with protein solutions freshly prepared before use in 5 mM phosphate buffer pH 7 and the protein concentration was checked spectrophotometrically, using e 405 = 121 700 M À1 cm À1 for both the M80A and M80A/Y67A mutants. 105,106…”

Section: Spectroscopic Measurementsmentioning

confidence: 99%

“…[1][2][3][4][5][6][7] The solution properties and binding events affect the heme environment and axial ligation, 1,[3][4][5][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27] modifying the physiological role in vivo and the reactivity in vitro. 1,5,9,[13][14][15][16]18,19,[28][29][30][31][32][33][34][35][36][37][38][39]…”

Section: Introductionmentioning

confidence: 99%

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Met80 and Tyr67 affect the chemical unfolding of yeast cytochromec: comparing the solutionvs.immobilized state

et al. 2020

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Section: Discussionmentioning

confidence: 99%

Section: Absorption and MCD Spectramentioning

confidence: 93%

Section: Spectroscopic Measurementsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Met80 and Tyr67 affect the chemical unfolding of yeast cytochromec: comparing the solutionvs.immobilized state

et al. 2020

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“…In vitro, there are many studies reporting on the interactions of cyt c with non-CL anionic lipids, including PG, showing the potential for similar interactions to those underpinning CL-induced peroxidase activation [28][29][30][31]. Yet, to the best of our knowledge there is no record of a direct involvement of PG in place of CL in the apoptotic pathway in vivo, with studies showing CL to be the preferred substrate for cyt c binding and peroxidation [2,3,32,33] [34]: the blue foldon (N and C-terminal helices, residues 1-14 and 88-104), the green foldon (helix [61][62][63][64][65][66][67][68][69] and Ω loop [20][21][22][23][24][25][26][27][28][29][30][31][32][33][34][35], the gray foldon (Ω loop [40][41][42][43][44][45][46][47][48][49][50][51]…”

Section: Introductionmentioning

confidence: 99%

Activation of Cytochrome C Peroxidase Function Through Coordinated Foldon Loop Dynamics upon Interaction with Anionic Lipids

Sun

Tyurin

et al. 2021

Preprint

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Cardiolipin (CL) is a mitochondrial anionic lipid that plays important roles in the regulation and signaling of mitochondrial apoptosis. CL peroxidation catalyzed by the assembly of CL-cytochrome c (cyt c) complexes at the inner mitochondrial membrane is a critical checkpoint. The structural changes in the protein, associated with peroxidase activation by CL and different anionic lipids, are not known at a molecular level. To better understand these peripheral protein-lipid interactions, we compare how phosphatidylglycerol (PG) and CL lipids trigger cyt c peroxidase activation, and correlate functional differences to structural and motional changes in membrane-associated cyt c. Structural and motional studies of the bound protein are enabled by magic angle spinning solid state NMR spectroscopy, while lipid peroxidase activity is assayed by mass spectrometry. PG binding results in a surface-bound state that preserves a nativelike fold, which nonetheless allows for significant peroxidase activity, though at a lower level than binding its native substrate CL. Lipid-specific differences in peroxidase activation are found to correlate to corresponding differences in lipid-induced protein mobility, affecting specific protein segments. The dynamics of omega loops C and D are upregulated by CL binding, in a way that is remarkably controlled by the protein:lipid stoichiometry. In contrast to complete chemical denaturation, membrane-induced protein destabilization reflects a destabilization of select cyt c foldons, while the energetically most stable helices are preserved. Our studies illuminate the interplay of protein and lipid dynamics in the creation of lipid peroxidase-active proteolipid complexes implicated in early stages of mitochondrial apoptosis.

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Electron Transfer and Electrocatalytic Properties of the Immobilized Met80Ala Cytochrome c Variant in Dimethylsulfoxide

et al. 2021

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The electrode-immobilized Met80Ala variant of yeast iso-1 cytochrome c in mixed water/dimethylsulfoxide (DMSO) solutions up to 60 % v/v DMSO shows thermodynamic and kinetic parameters of electron exchange and electrocatalytic properties towards O 2 reduction fully comparable to those in water. This is the result of moderate protein conformational changes thanks to immobilization that, to a certain extent, preserves protein structure, possibly due to the constraints on protein mobility/ flexibility induced by the electrostatic interactions with the electrode-coating SAM. Upon increasing the DMSO content of the mixed solution beyond 60 %, a much larger perturbation occurs that leads to the progressive loss of the electrocatalytic ability. Therefore, under these conditions, the organic solvent remarkably affects the structure and properties of the protein probably involving major conformational changes or even the replacement of the 6 th axial hydroxide ligand of the heme iron with a strong protein ligand, possibly a lysine residue.

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Binding of S. cerevisiae iso-1 cytochrome c and its surface lysine-to-alanine variants to cardiolipin: charge effects and the role of the lipid to protein ratio

Cited by 13 publications

References 104 publications

Met80 and Tyr67 affect the chemical unfolding of yeast cytochromec: comparing the solutionvs.immobilized state

Met80 and Tyr67 affect the chemical unfolding of yeast cytochromec: comparing the solutionvs.immobilized state

Activation of Cytochrome C Peroxidase Function Through Coordinated Foldon Loop Dynamics upon Interaction with Anionic Lipids

Electron Transfer and Electrocatalytic Properties of the Immobilized Met80Ala Cytochrome c Variant in Dimethylsulfoxide

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