A permselective-membrane electrode for the electrochemical study of redox proteins. Application to cytochrome c552 from Thiobacillus ferrooxidans

Haladjian, Jean; Bianco, Pierre; Nunzi, Frédéric; Bruschi, Mireille

doi:10.1016/0003-2670(94)80002-2

Cited by 62 publications

(52 citation statements)

References 5 publications

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“…At the very low pH of 2, the redox potential of Iro 33020 reached 550 mV. Note that the same evolution of redox potential with pH has been observed for rusticyanin, a periplasmic blue-copper protein (Haladjian et al, 1993), and cytochrome c 552 (Haladjian et al, 1994), from the same micro-organism.…”

supporting

confidence: 66%

“…Potentials versus the normal hydrogen electrode (NHE) have been obtained by adding 210 mV to the measured potentials (Bates, 1964). The membrane electrode technology was described in a previous paper (Haladjian et al, 1994). In such an electrode configuration, experiments require protein samples of only 1-2 ml, which were deposited on a square piece of dialysis membrane (Spectra/Por MWCO 6000-8000).…”

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confidence: 99%

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The HiPIP from the acidophilic Acidithiobacillus ferrooxidans is correctly processed and translocated in Escherichia coli, in spite of the periplasm pH difference between these two micro-organisms

et al. 2005

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The gene encoding a putative high-potential iron-sulfur protein (HiPIP) from the strictly acidophilic and chemolithoautotrophic Acidithiobacillus ferrooxidans ATCC 33020 has been cloned and sequenced. This potential HiPIP was overproduced in the periplasm of the neutrophile and heterotroph Escherichia coli. As shown by optical and EPR spectra and by electrochemical studies, the recombinant protein has all the biochemical properties of a HiPIP, indicating that the iron-sulfur cluster was correctly inserted. Translocation of this protein in the periplasm of E. coli was not detected in a DtatC mutant, indicating that it is dependent on the Tat system. The genetic organization of the iro locus in strains ATCC 23270 and ATCC 33020 is different from that found in strains Fe-1 and BRGM. Indeed, in A. ferrooxidans ATCC 33020 and ATCC 23270 (the type strain), iro was not located downstream from purA but was instead downstream from petC2, encoding cytochrome c 1 from the second A. ferrooxidans cytochrome bc 1 complex. These findings underline the genotypic heterogeneity within the A. ferrooxidans species. The results suggest that Iro transfers electrons from a cytochrome bc 1 complex to a terminal oxidase, as proposed for the HiPIP in photosynthetic bacteria. INTRODUCTIONOne of the most studied bacteria that thrives in acidic mine drainage is Acidithiobacillus ferrooxidans, formerly Thiobacillus ferrooxidans (Kelly & Wood, 2000;Leduc & Ferroni, 1994). It is characterized as a Gram-negative, acidophilic chemolithoautotroph that obtains energy for its growth mainly from the oxidation of ferrous iron (Fe 2+ ) or reduced sulfur compounds (Ingledew, 1982). Attempts to understand such an energetic metabolism, and especially the oxidation of Fe 2+ , have been initiated, leading to the proposition of several respiratory chains involving various redox proteins that have been identified and characterized in A. ferrooxidans (Fukumori et al., 1988;Blake & Shute, 1994;Yamanaka & Fukumori, 1995;Giudici-Orticoni et al., 1997Appia-Ayme, 1998;Appia-Ayme et al., 1999). Among them, a high-potential iron-sulfur protein (HiPIP) designated Iro, for iron-oxidizing enzyme, was characterized in A. ferrooxidans strains Fe-1 (Fukumori et al., 1988;Kusano et al., 1992) and BRGM (Cavazza et al., 1995) and the corresponding iro gene was also cloned and studied from strain Fe-1 (Kusano et al., 1992). The Iro protein was proposed to be the first electron acceptor in several alternative models of electron transfer chain between Fe 2+ and oxygen (Fukumori et al., 1988;Yamanaka et al., 1995). Based on genetic and subcellular localization studies with strain ATCC 33020, we proposed a model, not involving Iro, in which the electrons are passed from Fe 2+ to oxygen through a set of redox proteins expressed from an operon (Appia-Ayme, 1998;Appia-Ayme et al., 1998, 1999 Bengrine et al., 1998; Yarzábal et al., 2002) that is induced in Fe 2+ -grown cells (Yarzábal et al., 2004). To investigate the possible role of Iro, we probed for the presence of the iro Abbr...

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confidence: 66%

mentioning

confidence: 99%

The HiPIP from the acidophilic Acidithiobacillus ferrooxidans is correctly processed and translocated in Escherichia coli, in spite of the periplasm pH difference between these two micro-organisms

et al. 2005

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“…Thus, CDH IIB has an extended activity in the acidic and alkaline pH ranges. The pH optimum of CDH IIB for FcPF 6 is much higher than that of CDH IIA. The pH optima of various N. crassa cellulolytic enzymes range from pH 5.0 to 7.0, with significant residual activities from pH 3.0 to 8.5 (4,40).…”

Section: Discussionmentioning

confidence: 81%

“…Modification with CDH was done by filling the cavity with enzyme solution (20 mg ml Ϫ1 ). A dialysis membrane (molecular mass cutoff of 14,000 Da; Carl Roth) was used to trap the enzyme in the cells (6). The dialysis membrane (presoaked in buffer) was pressed onto the electrode and fixed tightly with a rubber O ring.…”

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confidence: 99%

Characterization of the Two Neurospora crassa Cellobiose Dehydrogenases and Their Connection to Oxidative Cellulose Degradation

Sygmund

Kracher

Scheiblbrandner

et al. 2012

Appl Environ Microbiol

117

125

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ABSTRACTThe genome ofNeurospora crassaencodes two different cellobiose dehydrogenases (CDHs) with a sequence identity of only 53%. So far, only CDH IIA, which is induced during growth on cellulose and features a C-terminal carbohydrate binding module (CBM), was detected in the secretome ofN. crassaand preliminarily characterized. CDH IIB is not significantly upregulated during growth on cellulosic material and lacks a CBM. Since CDH IIB could not be identified in the secretome, both CDHs were recombinantly produced inPichia pastoris. With the cytochrome domain-dependent one-electron acceptor cytochromec, CDH IIA has a narrower and more acidic pH optimum than CDH IIB. Interestingly, the catalytic efficiencies of both CDHs for carbohydrates are rather similar, but CDH IIA exhibits 4- to 5-times-higher apparent catalytic constants (kcatandKmvalues) than CDH IIB for most tested carbohydrates. A third major difference is the 65-mV-lower redox potential of the hemebcofactor in the cytochrome domain of CDH IIA than CDH IIB. To study the interaction with a member of the glycoside hydrolase 61 family, the copper-dependent polysaccharide monooxygenase GH61-3 (NCU02916) fromN. crassawas expressed inP. pastoris. A pH-dependent electron transfer from both CDHs via their cytochrome domains to GH61-3 was observed. The different properties of CDH IIA and CDH IIB and their effect on interactions with GH61-3 are discussed in regard to the proposedin vivofunction of the CDH/GH61 enzyme system in oxidative cellulose hydrolysis.

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“…The MEs were prepared as previously described [23,24]. A small volume (2 ml) of the protein solution was deposited on a square piece (about twice the diameter of the electrode sensor) of the dialysis membrane (Spectra/Por MWCO 6000 Á/8000, a negatively charged membrane, symbolized by M ( ), then the polished electrode was pressed against the membrane and a rubber ring was fitted around the electrode body so that the entrapped solution formed a uniform thin layer.…”

Section: Methodsmentioning

confidence: 99%

Electrochemical studies on small electron transfer proteins using membrane electrodes

Santos¹,

Sousa²,

Gonçalves³

et al. 2003

Journal of Electroanalytical Chemistry

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Membrane electrodes (ME) were constructed using gold, glassy carbon and pyrolytic graphite supports and a dialysis membrane, and used to study the electrochemical behavior of small size electron transfer proteins: monohemic cytochrome c 522 from Pseudomonas nautica and cytochrome c 533 as well as rubredoxin from Desulfovibrio vulgaris . Different electrochemical techniques were used including cyclic voltammetry (CV), square wave voltammetry (SW) and differential pulse voltammetry (DP). A direct electrochemical response was obtained in all cases except with rubredoxin where a facilitator was added to the protein solution entrapped between the membrane and the electrode surface. Formal potentials and heterogeneous charge transfer rate constants were determined from the voltammetric data. The influence of the ionic strength and the pH of the medium on the electrochemical response at the ME were analyzed. The benefits from the use of the ME in protein electrochemistry and its role in modulating the redox behavior are analyzed. A critical comparison is presented with data obtained at non-MEs. Finally, the interactions that must be established between the proteins and the electrode surfaces are discussed, thereby modeling molecular interactions that occur in biological systems. #

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A permselective-membrane electrode for the electrochemical study of redox proteins. Application to cytochrome c552 from Thiobacillus ferrooxidans

Cited by 62 publications

References 5 publications

The HiPIP from the acidophilic Acidithiobacillus ferrooxidans is correctly processed and translocated in Escherichia coli, in spite of the periplasm pH difference between these two micro-organisms

The HiPIP from the acidophilic Acidithiobacillus ferrooxidans is correctly processed and translocated in Escherichia coli, in spite of the periplasm pH difference between these two micro-organisms

Characterization of the Two Neurospora crassa Cellobiose Dehydrogenases and Their Connection to Oxidative Cellulose Degradation

Electrochemical studies on small electron transfer proteins using membrane electrodes

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