Bacterial Expression of a Mitochondrial Cytochromec. Trimethylation of Lys72 in Yeastiso-1-Cytochromecand the Alkaline Conformational Transition

Pollock, W. B. R.; Rosell, Federico I.; Twitchett, Mark B.; Dumont, and Mark E.; Mauk, A. Grant

doi:10.1021/bi972188d

Cited by 263 publications

(342 citation statements)

References 73 publications

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“…While stable isotope enrichment of cytochrome b 5 has been possible for several years, it is only recently that expression systems enabling stable isotope enrichment of cytochrome c have become available [16,28,53,54]. Consequently, we have taken advantage of this new possibility to obtain further insights on the present system, by monitoring the interaction from the point of view of both partners.…”

Section: Discussionmentioning

confidence: 99%

A further investigation of the cytochrome b 5–cytochrome c complex

et al. 2003

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The interaction of reduced rabbit cytochrome b 5 with reduced yeast iso-1 cytochrome c has been studied through the analysis of 1 H-15 N HSQC spectra, of 15 N longitudinal (R 1 ) and transverse (R 2 ) relaxation rates, and of the solvent exchange rates of protein backbone amides. For the first time, the adduct has been investigated also from the cytochrome c side. The analysis of the NMR data was integrated with docking calculations. The result is that cytochrome b 5 has two negative patches capable of interacting with a single positive surface area of cytochrome c. At low protein concentrations and in equimolar mixture, two different 1:1 adducts are formed. At high concentration and/or with excess cytochrome c, a 2:1 adduct is formed. All the species are in fast exchange on the scale of differences in chemical shift. By comparison with literature data, it appears that the structure of one 1:1 adduct changes with the origin or primary sequence of cytochrome b 5 .

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Section: Discussionmentioning

confidence: 99%

A further investigation of the cytochrome b 5–cytochrome c complex

et al. 2003

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“…Wild type recombinant untrimethylated C102T Saccharomyces cerevisiae iso-1-cytochrome c (ycc, hereafter) and the variant Lys72Ala/Lys73Ala/Lys79Ala (KtoA, hereafter) were expressed in E. coli and purified following the procedures described elsewhere [26,27,30], tyrosine replaces the cysteine at position 102 in both species. This substitution prevents dimerization and minimizes autoreduction while resulting in retention of the spectral and the functional properties of the protein [31,32].…”

Section: Methodsmentioning

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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“…Zn-substitutedCcP was prepared by heme extraction and reconstitution with Zn-protoporphyrin IX (Frontier Scientific) following the procedure of Asakura and Yonetani (33). The plasmid for yeast iso-1 Fe 3ϩ Cc(C102T) was obtained from Marcellus Ubbink (Leiden University, Leiden, The Netherlands), and Fe 3ϩ Cc was prepared as described (34).…”

Section: Methodsmentioning

confidence: 99%

Differential influence of dynamic processes on forward and reverse electron transfer across a protein-protein interface

Hoffman

Celis²,

Cull³

et al. 2005

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

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We propose that the forward and reverse halves of a flash-induced protein-protein electron transfer (ET) photocycle should exhibit differential responses to dynamic interconversion of configurations when the most stable configuration is not the most reactive, because the reactants exist in different initial configurations: the flash-photoinitiated forward ET process begins with the protein partners in an equilibrium ensemble of configurations, many of which have little or no reactivity, whereas the reactant of the thermal back ET (the charge-separated intermediate) is formed in a nonequilibrium, ''activated'' protein configuration. We report evidence for this proposal in measurements on (i) mixed-metal hemoglobin hybrids, (ii) the complex between cytochrome c peroxidase and cytochrome c, and (iii and iv) the complexes of myoglobin and isolated hemoglobin ␣-chains with cytochrome b 5. For all three systems, forward and reverse ET does respond differently to modulation of dynamic processes; further, the response to changes in viscosity is different for each system. cytochrome c ͉ dynamics ͉ hemoglobin ͉ myoglobin ͉ cytochrome c peroxidase T he long-range transfer of a single electron from donor to acceptor in a condensed phase is a fascinating and widely studied process (1, 2). Much of this work seeks to understand the electron transfer (ET) process itself. However, when the ET event involves a dynamic protein-protein interface, the observed kinetics frequently are controlled not by the ET process itself, but by the dynamics of recognition and binding and͞or conversion within an ensemble of bound configurations (3, 4).Studies of interprotein ET (3, 5-10) began with the implicit assumption of a protein-protein binding-energy landscape with a single reactive complex (Fig. 1A Left), implying a direct correlation between binding and reactivity. When the landscape for complex formation has several discrete minima ( Fig. 1 A Center) the reactive conformation may differ from the most stable one, in which case the observed ET kinetics are controlled by the rates and͞or energetics of conformational conversion within a complex (11-13). Recent studies of ET between myoglobin (Mb) and cytochrome b 5 (Fe 3ϩ b 5 ), which bind to each other by weak electrostatic interactions, disclosed a new dynamic docking paradigm in protein-protein reaction dynamics: the landscape involves numerous configurations of similar affinity, only a subset of which is active in ET (Fig. 1 A Right) (4, 14).A majority of these studies have used a photocycle in which laser-flash excitation of the metallo-porphyrin in a metalsubstituted (M ϭ Zn or Mg) hemoprotein to its triplet excited state ( 3 D) triggers ET from the triplet to the metal center of an acceptor protein (A) across a protein-protein interface, with rate constant, k f , Eq. 1:[1]The acceptor metal center of A typically is a ferri-heme center, Given the exponentially steep fall-off in the matrix element for ET between the two redox centers (19,20), there will be only a subset of conformation...

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Bacterial Expression of a Mitochondrial Cytochromec. Trimethylation of Lys72 in Yeastiso-1-Cytochromecand the Alkaline Conformational Transition

Cited by 263 publications

References 73 publications

A further investigation of the cytochrome b 5–cytochrome c complex

A further investigation of the cytochrome b 5–cytochrome c complex

Redox thermodynamics of cytochromes c subjected to urea induced unfolding

Differential influence of dynamic processes on forward and reverse electron transfer across a protein-protein interface

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