Peroxidase Activity as a Tool for Studying the Folding of<i>c</i>-Type Cytochromes

Diederix, Rutger E. M.; Ubbink, Marcellus; Canters, Gerard W.

doi:10.1021/bi0260841

Cited by 108 publications

(141 citation statements)

References 46 publications

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“…Peroxidase activity was assayed with guaiacol (Sigma) by measuring absorption at 470 nm of the formed tetraguaiacol product (48). Cyt c preincubated in a liposome solution for at least 2 h was added to a cuvette containing a freshly prepared mixture of guaiacol and hydrogen peroxide, H 2 O 2 .…”

Section: Preparation Of Lipid Vesicles and Protein-liposome Binding Ementioning

confidence: 99%

Conformational properties of cardiolipin-bound cytochrome c

Hanske

Toffey

Morenz

et al. 2011

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

189

388

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Interactions of cytochrome c (cyt c) with cardiolipin (CL) are important for both electron transfer and apoptotic functions of this protein. A sluggish peroxidase in its native state, when bound to CL, cyt c catalyzes CL peroxidation, which contributes to the protein apoptotic release. The heterogeneous CL-bound cyt c ensemble is difficult to characterize with traditional structural methods and ensemble-averaged probes. We have employed time-resolved FRET measurements to evaluate structural properties of the CL-bound protein in four dansyl (Dns)-labeled variants of horse heart cyt c. The Dns decay curves and extracted Dns-to-heme distance distributions PðrÞ reveal a conformational diversity of the CL-bound cyt c ensemble with distinct populations of the polypeptide structures that vary in their degree of protein unfolding. A fraction of the ensemble is substantially unfolded, with Dns-to-heme distances resembling those in the guanidine hydrochloride-denatured state. These largely open cyt c structures likely dominate the peroxidase activity of the CL-bound cyt c ensemble. Site variations in PðrÞ distributions uncover structural features of the CL-bound cyt c, rationalize previous findings, and implicate the prime role of electrostatic interactions, particularly with the protein C terminus, in the CL-induced unfolding.membrane | redox protein | fluorescence

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Section: Preparation Of Lipid Vesicles and Protein-liposome Binding Ementioning

confidence: 99%

Conformational properties of cardiolipin-bound cytochrome c

Hanske

Toffey

Morenz

et al. 2011

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

189

388

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“…Although the unfolding effect of urea on cytochromes c is discussed at a fundamental level, the implications of this systems in biocatalysis are worth considering. In fact, the peroxidase activity of cytochrome c increases substantially upon unfolding [28,29].…”

Section: Introductionmentioning

confidence: 99%

Redox thermodynamics of cytochromes c subjected to urea induced unfolding

et al. 2009

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The thermodynamics of the electron transfer (ET) process for beef heart and yeast cytochromes c and the Lys72Ala/Lys73Ala/Lys79Ala mutant of the latter species subjected to progressive urea-induced unfolding was determined electrochemically. The results indicate the presence of at least three protein forms which were assigned to a low-temperature and a high-temperature HisMet intermediate species and a bis-histidinate form (although the presence of a His-Lys form cannot be excluded). The much lower E°0 value of the bis-histidinate conformer as compared to His-Met ligated species is largely determined by the enthalpic contribution induced by axial ligand substitution. The biphasic E°0 versus T profile for the His-Met species is due to a difference in reduction entropy between the conformers at low and high temperatures. Enthalpy-entropy compensation phenomena for the reduction reaction at varying urea concentration for all the forms of the investigated cytochromes c were addressed and discussed.

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“…Previous studies have proven that native cyt c does not contain any peroxidase activity; however, very small structural variation of cyt c that cannot be readily probed in spectroscopy can lead to significant increase in the peroxidase activity of cyt c towards the catalytic reduction of hydrogen peroxidase (H 2 O 2 ). Thus the peroxidase activity of cyt c provides an extremely sensitive probe for the structural intactness of cyt c. 56 Figure 5e shows bio-electrocatalytic response of cyt c towards 10 µM H 2 O 2 in Na-PBS (pH 7.4). It shows that with H 2 O 2 concentration, the reduction peak current is obviously increased, while the oxidation peak current decreased, which is typical electrocatalytic reduction process of H 2 O 2 .…”

Section: Protein 1: Cytochrome C (Cyt C)mentioning

confidence: 99%

Probing the nature of electron transfer in metalloproteins on graphene-family materials as nanobiocatalytic scaffold using electrochemistry

Gupta

Irihamye

2015

AIP Advances

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Graphene-based nanomaterials have shown great promise not only in nanoelectronics due to ultrahigh electron mobility but also as biocatalytic scaffolds owing to irreversible protein surface adsorption and facilitating direct electron transfer. In this work, we synthesized stable dispersions of graphene using liquid-phase exfoliation approach based on non-covalent interactions between graphene and 1-pyrenesulfonic acid sodium salt (Py–1SO3), 1-pyrenemethylamine salt (Py − Me-NH2) and Pluronic® P-123 surfactant using only water as solvent compatible with biomolecules. The resulting graphene nanoplatelets (Gr_LPE) are characterized by a combination of analytical (microscopy and spectroscopy) techniques revealing mono- to few-layer graphene displaying that the exfoliation efficiency strongly depends upon the type of pyrene-based salts and organic surfactants. Moreover being completely water-based approach, we build robust nanoscaffolds of graphene-family nanomaterials (GFNs) namely, monolayer graphene, Gr_LPE (the one prepared with Pluronic® P-123), graphene oxide (GO) and its reduced form (rGO) on glassy carbon electrode surface with three important metalloproteins include cytochrome c (Cyt c) [for electron transfer], myoglobin (Mb) [for oxygen storage] and horseradish peroxidase (HRP) [for catalyzing the biochemical reaction]. In order to demonstrate the nanobiocatalytical activity of these proteins, we used electrochemical interfacial direct electron transfer (DET) kinetics and attempt to determine the rate constant (kET) using two different analytical approaches namely, linear sweep voltammetry and Laviron’s theory. We elucidated that all of the metalloproteins retain their structural integrity (secondary structure) upon forming mixtures with GFNs confirmed through optical and vibrational spectroscopy and biological activity using electrochemistry. Among the GFNs studied, Gr-LPE, GO and rGO support the efficient electrical wiring of the redox centers (with an increase in catalytic efficiency of Cyt c and Mb in the presence of GFNs attributed partially to the surface functional (carboxyl, epoxide and hydroxyl) groups on GO and rGO facilitating rapid charge transfer.

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Peroxidase Activity as a Tool for Studying the Folding ofc-Type Cytochromes

Cited by 108 publications

References 46 publications

Conformational properties of cardiolipin-bound cytochrome c

Conformational properties of cardiolipin-bound cytochrome c

Redox thermodynamics of cytochromes c subjected to urea induced unfolding

Probing the nature of electron transfer in metalloproteins on graphene-family materials as nanobiocatalytic scaffold using electrochemistry

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