Efficient and Selective Isotopic Labeling of Hemes to Facilitate the Study of Multiheme Proteins

Fonseca, Bruno M.; Rivera, Mario; Shi, Liang; Louro, Ricardo O.

doi:10.2144/000113859

Cited by 10 publications

(7 citation statements)

References 51 publications

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“…This rendered the isotopic labeling of multiheme cytochromes much more cost-effective (Fernandes et al, 2008 ), facilitated the NMR signal assignment procedure and provided the foundations to identify redox partners and map their interacting regions (Dantas et al, 2014 ). A methodology that enables the isotopic labeling of multiheme cytochromes exclusively in their redox centers was recently reported (Fonseca et al, 2012b ) and is expected to be a valuable tool in the assignment of heme signals in very large multiheme cytochromes for which the overlap of signals in the NMR spectra is severe.…”

Section: Methodological Improvements To Assist the Solution Structuramentioning

confidence: 99%

Rational engineering of Geobacter sulfurreducens electron transfer components: a foundation for building improved Geobacter-based bioelectrochemical technologies

et al. 2015

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Multiheme cytochromes have been implicated in Geobacter sulfurreducens extracellular electron transfer (EET). These proteins are potential targets to improve EET and enhance bioremediation and electrical current production by G. sulfurreducens. However, the functional characterization of multiheme cytochromes is particularly complex due to the co-existence of several microstates in solution, connecting the fully reduced and fully oxidized states. Over the last decade, new strategies have been developed to characterize multiheme redox proteins functionally and structurally. These strategies were used to reveal the functional mechanism of G. sulfurreducens multiheme cytochromes and also to identify key residues in these proteins for EET. In previous studies, we set the foundations for enhancement of the EET abilities of G. sulfurreducens by characterizing a family of five triheme cytochromes (PpcA-E). These periplasmic cytochromes are implicated in electron transfer between the oxidative reactions of metabolism in the cytoplasm and the reduction of extracellular terminal electron acceptors at the cell's outer surface. The results obtained suggested that PpcA can couple e−/H+ transfer, a property that might contribute to the proton electrochemical gradient across the cytoplasmic membrane for metabolic energy production. The structural and functional properties of PpcA were characterized in detail and used for rational design of a family of 23 single site PpcA mutants. In this review, we summarize the functional characterization of the native and mutant proteins. Mutants that retain the mechanistic features of PpcA and adopt preferential e−/H+ transfer pathways at lower reduction potential values compared to the wild-type protein were selected for in vivo studies as the best candidates to increase the electron transfer rate of G. sulfurreducens. For the first time G. sulfurreducens strains have been manipulated by the introduction of mutant forms of essential proteins with the aim to develop and improve bioelectrochemical technologies.

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Section: Methodological Improvements To Assist the Solution Structuramentioning

confidence: 99%

Rational engineering of Geobacter sulfurreducens electron transfer components: a foundation for building improved Geobacter-based bioelectrochemical technologies

et al. 2015

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“…Magnetic fields was reported to cause stress in bacterial cells and activates genes ALA dehydratase, which is a key enzyme from a synthesis porphyrin, and subsequently increase the production of endogenous photosensitizing porphyrin (Fonseca et al, 2012). As a result, there might be a synergistic effect between aBL and magnetic fields.…”

Section: Synergism Of Antimicrobial Blue Light With Other Agentsmentioning

confidence: 99%

Antimicrobial blue light inactivation of pathogenic microbes: State of the art

Wang

et al. 2017

Drug Resistance Updates

225

192

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As an innovative non-antibiotic approach, antimicrobial blue light in the spectrum of 400–470 nm has demonstrated its intrinsic antimicrobial properties resulting from the presence of endogenous photosensitizing chromophores in pathogenic microbes and, subsequently, its promise as a counteracter of antibiotic resistance. Since we published our last review of antimicrobial blue light in 2012, there have been a substantial number of new studies reported in this area. Here we provide an updated overview of the findings from the new studies over the past 5 years, including the efficacy of antimicrobial blue light inactivation of different microbes, its mechanism of action, synergism of antimicrobial blue light with other angents, its effect on host cells and tissues, the potential development of resistance to antimicrobial blue light by microbes, and a novel interstitial delivery approach of antimicrobial blue light. The potential new applications of antimicrobial blue light are also discussed.

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“…Hence, they might also be deployed to experimentally deconvolute voltammetric responses in terms of individual cofactors (by observing the impact of a given haem modification on the voltammogram). Also, the NMR methods to determine microscopic redox potentials of individual haems could possibly be extended to larger cytochromes as well in combination with 13 C-labelling of haem methyl groups [84,157].…”

Section: Experimental Approaches To Intraprotein Electron Transportmentioning

confidence: 99%

Multi-haem cytochromes inShewanella oneidensisMR-1: structures, functions and opportunities

Breuer

Rosso

Blumberger

et al. 2015

J. R. Soc. Interface.

220

186

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Multi-haem cytochromes are employed by a range of microorganisms to transport electrons over distances of up to tens of nanometres. Perhaps the most spectacular utilization of these proteins is in the reduction of extracellular solid substrates, including electrodes and insoluble mineral oxides of Fe(III) and Mn(III/IV), by species of Shewanella and Geobacter. However, multihaem cytochromes are found in numerous and phylogenetically diverse prokaryotes where they participate in electron transfer and redox catalysis that contributes to biogeochemical cycling of N, S and Fe on the global scale. These properties of multi-haem cytochromes have attracted much interest and contributed to advances in bioenergy applications and bioremediation of contaminated soils. Looking forward, there are opportunities to engage multi-haem cytochromes for biological photovoltaic cells, microbial electrosynthesis and developing bespoke molecular devices. As a consequence, it is timely to review our present understanding of these proteins and we do this here with a focus on the multitude of functionally diverse multi-haem cytochromes in Shewanella oneidensis MR-1. We draw on findings from experimental and computational approaches which ideally complement each other in the study of these systems: computational methods can interpret experimentally determined properties in terms of molecular structure to cast light on the relation between structure and function. We show how this synergy has contributed to our understanding of multi-haem cytochromes and can be expected to continue to do so for greater insight into natural processes and their informed exploitation in biotechnologies.

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Efficient and Selective Isotopic Labeling of Hemes to Facilitate the Study of Multiheme Proteins

Cited by 10 publications

References 51 publications

Rational engineering of Geobacter sulfurreducens electron transfer components: a foundation for building improved Geobacter-based bioelectrochemical technologies

Rational engineering of Geobacter sulfurreducens electron transfer components: a foundation for building improved Geobacter-based bioelectrochemical technologies

Antimicrobial blue light inactivation of pathogenic microbes: State of the art

Multi-haem cytochromes inShewanella oneidensisMR-1: structures, functions and opportunities

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