Filipa Calisto - Academia.edu (original) (raw)

Papers by Filipa Calisto

"Life depends on a constant energy supply and conversion. Cells use energy from external sou... more "Life depends on a constant energy supply and conversion. Cells use energy from external sources, such as organic or inorganic compounds or light to sustain, grow and reproduce themselves. The energy from those sources has to be converted into energy forms useable for the different cellular processes, i.e., external energy is converted (transduced) to other forms such as adenosine triphosphate (ATP) and/or a transmembrane difference of the electrochemical potential.(...)

Frontiers in Chemistry, 2021

Several energy-transducing microbial enzymes have their peripheral subunits connected to the memb... more Several energy-transducing microbial enzymes have their peripheral subunits connected to the membrane through an integral membrane protein, that interacts with quinones but does not have redox cofactors, the so-called NrfD-like subunit. The periplasmic nitrite reductase (NrfABCD) was the first complex recognized to have a membrane subunit with these characteristics and consequently provided the family's name: NrfD. Sequence analyses indicate that NrfD homologs are present in many diverse enzymes, such as polysulfide reductase (PsrABC), respiratory alternative complex III (ACIII), dimethyl sulfoxide (DMSO) reductase (DmsABC), tetrathionate reductase (TtrABC), sulfur reductase complex (SreABC), sulfite dehydrogenase (SoeABC), quinone reductase complex (QrcABCD), nine-heme cytochrome complex (NhcABCD), group-2 [NiFe] hydrogenase (Hyd-2), dissimilatory sulfite-reductase complex (DsrMKJOP), arsenate reductase (ArrC) and multiheme cytochrome c sulfite reductase (MccACD). The molecular...

Biochemical Society Transactions, 2021

Energy transduction is the conversion of one form of energy into another; this makes life possibl... more Energy transduction is the conversion of one form of energy into another; this makes life possible as we know it. Organisms have developed different systems for acquiring energy and storing it in useable forms: the so-called energy currencies. A universal energy currency is the transmembrane difference of electrochemical potential (Δμ~). This results from the translocation of charges across a membrane, powered by exergonic reactions. Different reactions may be coupled to charge-translocation and, in the majority of cases, these reactions are catalyzed by modular enzymes that always include a transmembrane subunit. The modular arrangement of these enzymes allows for different catalytic and charge-translocating modules to be combined. Thus, a transmembrane charge-translocating module can be associated with different catalytic subunits to form an energy-transducing complex. Likewise, the same catalytic subunit may be combined with a different membrane charge-translocating module. In th...

Advances in Microbial Physiology, 2019

The diversity of microbial cells is reflected in differences in cell size and shape, motility, me... more The diversity of microbial cells is reflected in differences in cell size and shape, motility, mechanisms of cell division, pathogenicity or adaptation to different environmental niches. All these variations are achieved by the distinct metabolic strategies adopted by the organisms. The respiratory chains are integral parts of those strategies especially because they perform the most or, at least, most efficient energy conservation in the cell. Respiratory chains are composed of several membrane proteins, which perform a stepwise oxidation of metabolites toward the reduction of terminal electron acceptors. Many of these membrane proteins use the energy released from the oxidoreduction reaction they catalyze to translocate charges across the membrane and thus contribute to the establishment of the membrane potential, i.e. they conserve energy. In this work we illustrate and discuss the composition of the respiratory chains of different taxonomic clades, based on bioinformatic analyses and on biochemical data available in the literature. We explore the diversity of the respiratory chains of Animals, Plants, Fungi and Protists kingdoms as well as of Prokaryotes, including Bacteria and Archaea. The prokaryotic phyla studied in this work are Gammaproteobacteria, Betaproteobacteria, Epsilonproteobacteria, Deltaproteobacteria, Alphaproteobacteria, Firmicutes, Actinobacteria, Chlamydiae, Verrucomicrobia, Acidobacteria, Planctomycetes, Cyanobacteria, Bacteroidetes, Chloroflexi, Deinococcus-Thermus, Aquificae, Thermotogae, Deferribacteres, Nitrospirae, Euryarchaeota, Crenarchaeota and Thaumarchaeota.

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2018

Tasas de interés de los fondos del Sistema de Banca para el Desarrollo y del sistema financiero, ... more Tasas de interés de los fondos del Sistema de Banca para el Desarrollo y del sistema financiero, 2016 ...

Antibiotics, 2018

Herein, we describe a case report of carbapenem-resistant Acinetobacter baumannii and Klebsiella ... more Herein, we describe a case report of carbapenem-resistant Acinetobacter baumannii and Klebsiella pneumoniae isolates that were identified from the same patient at a Tertiary University Hospital Centre in Portugal. Antimicrobial susceptibility and the molecular characterization of resistance and virulence determinants were performed. PCR screening identified the presence of the resistance genes blaKPC-3, blaTEM-1 and blaSHV-1 in both isolates. The KPC-3 K. pneumoniae isolate belonged to the ST-14 high risk clone and accumulated an uncommon resistance and virulence profile additional to a horizontal dissemination capacity. In conclusion, the molecular screening led to the first identification of the A. baumannii KPC-3 producer in Portugal with a full antimicrobial resistance profile including tigecycline and colistin.

Nature communications, Jan 30, 2018

Electron transfer in respiratory chains generates the electrochemical potential that serves as en... more Electron transfer in respiratory chains generates the electrochemical potential that serves as energy source for the cell. Prokaryotes can use a wide range of electron donors and acceptors and may have alternative complexes performing the same catalytic reactions as the mitochondrial complexes. This is the case for the alternative complex III (ACIII), a quinol:cytochrome c/HiPIP oxidoreductase. In order to understand the catalytic mechanism of this respiratory enzyme, we determined the structure of ACIII from Rhodothermus marinus at 3.9 Å resolution by single-particle cryo-electron microscopy. ACIII presents a so-far unique structure, for which we establish the arrangement of the cofactors (four iron-sulfur clusters and six c-type hemes) and propose the location of the quinol-binding site and the presence of two putative proton pathways in the membrane. Altogether, this structure provides insights into a mechanism for energy transduction and introduces ACIII as a redox-driven proton...

Biological Chemistry, 2017

Alternative Complex III (ACIII) is an example of the robustness and flexibility of prokaryotic re... more Alternative Complex III (ACIII) is an example of the robustness and flexibility of prokaryotic respiratory chains. It performs quinol:cytochrome

Biochimica et biophysica acta, 2016

Acquisition of energy is central to life. In addition to the synthesis of ATP, organisms need ene... more Acquisition of energy is central to life. In addition to the synthesis of ATP, organisms need energy for the establishment and maintenance of a transmembrane difference in electrochemical potential allowing cells to import and export metabolites or to their motility. The membrane potential is established by a variety of membrane bound respiratory complexes. In this work we explored the diversity of membrane respiratory chains and the presence of the different enzyme complexes in the several phyla of life. We performed taxonomic profiles of the several membrane bound respiratory proteins and complexes evaluating the presence of their respective coding genes in all species deposited in KEGG database. We evaluated 26 quinone reductases, 5 quinol:electron carriers oxidoreductases and 18 terminal electron acceptor reductases. We further included in the analyses enzymes performing redox or decarboxylation driven ion translocation, ATP synthase and transhydrogenase and we also investigated...

We conducted a surveillance study of carbapenem­resistant Enterobacteriaceae isolates in order to... more We conducted a surveillance study of carbapenem­resistant Enterobacteriaceae isolates in order to investigate the KPC­3 carbapenemase dissemination in Portugal. Methods: A PCR­based, hospital surveillance study of carbapenem resistance in Enterobacteriacea isolates was conducted in Portugal from January 2010 to July 2011 in a tertiary care Group of Hospitals, in Lisbon. Antimicrobial susceptibility testing was performed using disk diffusion method and interpreted following the CLSI. The presence of genes encoding metallo­beta­lactamases (IMP, VIM, NDM), KPC­type and OXA­48 and others ESBL enzymes were screened by PCR method using specific primers. All amplified products were then subjected to direct nucleotide sequencing. Results: Since the first description in Portugal in 2009, 43 unique and multi­resistant isolates were identified. 48% of the isolates were from surgical wards, 21% from intensive and intermediary care units and 9% from surgical outpatients follow­up. The KPC­3 carb...

The emergence and global spread of Enterobacteriaceae carbapenem-resistant is a public health thr... more The emergence and global spread of Enterobacteriaceae carbapenem-resistant is a public health threat because they are associated with high morbidity and mortality. The aim of this study was to characterize the blaKPC-3 gene, their genetic environment and other resistances associated in bacteria with reduced susceptibility to carbapenems in the North of Lisbon Hospital Centre, EPE. Forty-one clinical isolates of Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Escherichia coli and Citrobacter freundii were identified between November 2009 and June 2011 and all isolates were KPC-3 carbapenemase producing. Associated with this carbapenemase we detected other ß-lactamases: broad spectrum ß-lactamases, TEM-1 and SHV-1; extended-spectrum ß-lactamases SHV-12, SHV-35, CTX-M-15 and DHA-1 cefalosporinase. The absence of OmpK35 and OmpK36 outer membrane porins justify the increased of resistance in the strains to carbapenems and cephalosporins. The gene blaKPC-3 is associated...

Objectives: Nowadays the carbapenems resistance among Acinetobacter baumannii and Klebsiella pneu... more Objectives: Nowadays the carbapenems resistance among Acinetobacter baumannii and Klebsiella pneumoniae are serious therapeutic and infection control challenge. The aim of this study was to analyse the clinical characteristics and to investigate the genetic basis of the carbapenem resistance of K. pneumoniae and A. baumannii clinical strains isolated from the same patient. Methods: Antimicrobial susceptibility was determined by disk diffusion (amoxicillin/clavulanic acid, cefotaxime, cefoxitine, ceftazidime, imipenem, meropenem, ciprofloxacin, gentamicin, tigecycline and colistin), and interpreted according to CLSI guidelines. The presence of blaTEM, blaSHV, blaCTX, blaKPC, blaIMP and blaVIM was screened by PCR using specific primers. Amplicons were sequenced and compared to sequences available in the GeneBank database. Results: A female patient, with 35 years old, had renal insufficiency, being an immunocompromised host. She had been submitted to ciprofloxacin and meropenem therapy...

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2014

under anaerobic conditions and it enables anaerobic growth with urocanic acid as a sole terminal ... more under anaerobic conditions and it enables anaerobic growth with urocanic acid as a sole terminal electron acceptor. The latter's capability can provide the cells of UrdA-containing bacteria with a niche where no other bacteria can compete and survive.

ABSTRACT Respiratory chains are composed of several membrane protein complexes enabling the trans... more ABSTRACT Respiratory chains are composed of several membrane protein complexes enabling the transduction of energy from oxidoreduction reactions to the establishment of transmembrene difference of the membrane potential by charge translocation. Central to aerobic respiratory chains is cytochrome bc1 complex, which presents quinol:cytochrome c oxidoreductase activity. We recognize the existence of an alternative complex , Alternative Complex III – ACIII, performing the same function as the bc 1 complex but structurally totally distinct. Besides four transmembrene proteins, ACIII contains three peripheral subunits, ActA and ActE are facing the periplasm but the orientation in relation to the membrane of the ActB remains unknown. In this work we investigated the orientation of subunit ActB and its functional implications, taking advantage of ProteinaseK activity on isolated membrane vesicles from R. marinus.

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2013

The increasing number of completely sequenced genomes led us to perform a new search for the gene... more The increasing number of completely sequenced genomes led us to perform a new search for the genes coding Available online 9 January 2013 for the different ACIII subunits. We have identified a larger number of gene clusters coding for ACIII, still confined to the bacterial domain, but extended to classes in which it was not observed before. We also found an unantic-Keywords: Alternative complex III ipated diversity in gene clusters, both in terms of its constitution and organization. The several unexpected gene CISM family arrangements brought new perspectives to the role of the different subunits of ACIII, namely in quinone binding Heme copper oxygen reductase and in proton translocation. This article is part of a Special Issue entitled: Respiratory complex III and related bc Rhodothermus marinus complexes.

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2016

"Life depends on a constant energy supply and conversion. Cells use energy from external sou... more "Life depends on a constant energy supply and conversion. Cells use energy from external sources, such as organic or inorganic compounds or light to sustain, grow and reproduce themselves. The energy from those sources has to be converted into energy forms useable for the different cellular processes, i.e., external energy is converted (transduced) to other forms such as adenosine triphosphate (ATP) and/or a transmembrane difference of the electrochemical potential.(...)

Frontiers in Chemistry, 2021

Several energy-transducing microbial enzymes have their peripheral subunits connected to the memb... more Several energy-transducing microbial enzymes have their peripheral subunits connected to the membrane through an integral membrane protein, that interacts with quinones but does not have redox cofactors, the so-called NrfD-like subunit. The periplasmic nitrite reductase (NrfABCD) was the first complex recognized to have a membrane subunit with these characteristics and consequently provided the family's name: NrfD. Sequence analyses indicate that NrfD homologs are present in many diverse enzymes, such as polysulfide reductase (PsrABC), respiratory alternative complex III (ACIII), dimethyl sulfoxide (DMSO) reductase (DmsABC), tetrathionate reductase (TtrABC), sulfur reductase complex (SreABC), sulfite dehydrogenase (SoeABC), quinone reductase complex (QrcABCD), nine-heme cytochrome complex (NhcABCD), group-2 [NiFe] hydrogenase (Hyd-2), dissimilatory sulfite-reductase complex (DsrMKJOP), arsenate reductase (ArrC) and multiheme cytochrome c sulfite reductase (MccACD). The molecular...

Biochemical Society Transactions, 2021

Energy transduction is the conversion of one form of energy into another; this makes life possibl... more Energy transduction is the conversion of one form of energy into another; this makes life possible as we know it. Organisms have developed different systems for acquiring energy and storing it in useable forms: the so-called energy currencies. A universal energy currency is the transmembrane difference of electrochemical potential (Δμ~). This results from the translocation of charges across a membrane, powered by exergonic reactions. Different reactions may be coupled to charge-translocation and, in the majority of cases, these reactions are catalyzed by modular enzymes that always include a transmembrane subunit. The modular arrangement of these enzymes allows for different catalytic and charge-translocating modules to be combined. Thus, a transmembrane charge-translocating module can be associated with different catalytic subunits to form an energy-transducing complex. Likewise, the same catalytic subunit may be combined with a different membrane charge-translocating module. In th...

Advances in Microbial Physiology, 2019

The diversity of microbial cells is reflected in differences in cell size and shape, motility, me... more The diversity of microbial cells is reflected in differences in cell size and shape, motility, mechanisms of cell division, pathogenicity or adaptation to different environmental niches. All these variations are achieved by the distinct metabolic strategies adopted by the organisms. The respiratory chains are integral parts of those strategies especially because they perform the most or, at least, most efficient energy conservation in the cell. Respiratory chains are composed of several membrane proteins, which perform a stepwise oxidation of metabolites toward the reduction of terminal electron acceptors. Many of these membrane proteins use the energy released from the oxidoreduction reaction they catalyze to translocate charges across the membrane and thus contribute to the establishment of the membrane potential, i.e. they conserve energy. In this work we illustrate and discuss the composition of the respiratory chains of different taxonomic clades, based on bioinformatic analyses and on biochemical data available in the literature. We explore the diversity of the respiratory chains of Animals, Plants, Fungi and Protists kingdoms as well as of Prokaryotes, including Bacteria and Archaea. The prokaryotic phyla studied in this work are Gammaproteobacteria, Betaproteobacteria, Epsilonproteobacteria, Deltaproteobacteria, Alphaproteobacteria, Firmicutes, Actinobacteria, Chlamydiae, Verrucomicrobia, Acidobacteria, Planctomycetes, Cyanobacteria, Bacteroidetes, Chloroflexi, Deinococcus-Thermus, Aquificae, Thermotogae, Deferribacteres, Nitrospirae, Euryarchaeota, Crenarchaeota and Thaumarchaeota.

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2018

Tasas de interés de los fondos del Sistema de Banca para el Desarrollo y del sistema financiero, ... more Tasas de interés de los fondos del Sistema de Banca para el Desarrollo y del sistema financiero, 2016 ...

Antibiotics, 2018

Herein, we describe a case report of carbapenem-resistant Acinetobacter baumannii and Klebsiella ... more Herein, we describe a case report of carbapenem-resistant Acinetobacter baumannii and Klebsiella pneumoniae isolates that were identified from the same patient at a Tertiary University Hospital Centre in Portugal. Antimicrobial susceptibility and the molecular characterization of resistance and virulence determinants were performed. PCR screening identified the presence of the resistance genes blaKPC-3, blaTEM-1 and blaSHV-1 in both isolates. The KPC-3 K. pneumoniae isolate belonged to the ST-14 high risk clone and accumulated an uncommon resistance and virulence profile additional to a horizontal dissemination capacity. In conclusion, the molecular screening led to the first identification of the A. baumannii KPC-3 producer in Portugal with a full antimicrobial resistance profile including tigecycline and colistin.

Nature communications, Jan 30, 2018

Electron transfer in respiratory chains generates the electrochemical potential that serves as en... more Electron transfer in respiratory chains generates the electrochemical potential that serves as energy source for the cell. Prokaryotes can use a wide range of electron donors and acceptors and may have alternative complexes performing the same catalytic reactions as the mitochondrial complexes. This is the case for the alternative complex III (ACIII), a quinol:cytochrome c/HiPIP oxidoreductase. In order to understand the catalytic mechanism of this respiratory enzyme, we determined the structure of ACIII from Rhodothermus marinus at 3.9 Å resolution by single-particle cryo-electron microscopy. ACIII presents a so-far unique structure, for which we establish the arrangement of the cofactors (four iron-sulfur clusters and six c-type hemes) and propose the location of the quinol-binding site and the presence of two putative proton pathways in the membrane. Altogether, this structure provides insights into a mechanism for energy transduction and introduces ACIII as a redox-driven proton...

Biological Chemistry, 2017

Alternative Complex III (ACIII) is an example of the robustness and flexibility of prokaryotic re... more Alternative Complex III (ACIII) is an example of the robustness and flexibility of prokaryotic respiratory chains. It performs quinol:cytochrome

Biochimica et biophysica acta, 2016

Acquisition of energy is central to life. In addition to the synthesis of ATP, organisms need ene... more Acquisition of energy is central to life. In addition to the synthesis of ATP, organisms need energy for the establishment and maintenance of a transmembrane difference in electrochemical potential allowing cells to import and export metabolites or to their motility. The membrane potential is established by a variety of membrane bound respiratory complexes. In this work we explored the diversity of membrane respiratory chains and the presence of the different enzyme complexes in the several phyla of life. We performed taxonomic profiles of the several membrane bound respiratory proteins and complexes evaluating the presence of their respective coding genes in all species deposited in KEGG database. We evaluated 26 quinone reductases, 5 quinol:electron carriers oxidoreductases and 18 terminal electron acceptor reductases. We further included in the analyses enzymes performing redox or decarboxylation driven ion translocation, ATP synthase and transhydrogenase and we also investigated...

We conducted a surveillance study of carbapenem­resistant Enterobacteriaceae isolates in order to... more We conducted a surveillance study of carbapenem­resistant Enterobacteriaceae isolates in order to investigate the KPC­3 carbapenemase dissemination in Portugal. Methods: A PCR­based, hospital surveillance study of carbapenem resistance in Enterobacteriacea isolates was conducted in Portugal from January 2010 to July 2011 in a tertiary care Group of Hospitals, in Lisbon. Antimicrobial susceptibility testing was performed using disk diffusion method and interpreted following the CLSI. The presence of genes encoding metallo­beta­lactamases (IMP, VIM, NDM), KPC­type and OXA­48 and others ESBL enzymes were screened by PCR method using specific primers. All amplified products were then subjected to direct nucleotide sequencing. Results: Since the first description in Portugal in 2009, 43 unique and multi­resistant isolates were identified. 48% of the isolates were from surgical wards, 21% from intensive and intermediary care units and 9% from surgical outpatients follow­up. The KPC­3 carb...

The emergence and global spread of Enterobacteriaceae carbapenem-resistant is a public health thr... more The emergence and global spread of Enterobacteriaceae carbapenem-resistant is a public health threat because they are associated with high morbidity and mortality. The aim of this study was to characterize the blaKPC-3 gene, their genetic environment and other resistances associated in bacteria with reduced susceptibility to carbapenems in the North of Lisbon Hospital Centre, EPE. Forty-one clinical isolates of Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter aerogenes, Escherichia coli and Citrobacter freundii were identified between November 2009 and June 2011 and all isolates were KPC-3 carbapenemase producing. Associated with this carbapenemase we detected other ß-lactamases: broad spectrum ß-lactamases, TEM-1 and SHV-1; extended-spectrum ß-lactamases SHV-12, SHV-35, CTX-M-15 and DHA-1 cefalosporinase. The absence of OmpK35 and OmpK36 outer membrane porins justify the increased of resistance in the strains to carbapenems and cephalosporins. The gene blaKPC-3 is associated...

Objectives: Nowadays the carbapenems resistance among Acinetobacter baumannii and Klebsiella pneu... more Objectives: Nowadays the carbapenems resistance among Acinetobacter baumannii and Klebsiella pneumoniae are serious therapeutic and infection control challenge. The aim of this study was to analyse the clinical characteristics and to investigate the genetic basis of the carbapenem resistance of K. pneumoniae and A. baumannii clinical strains isolated from the same patient. Methods: Antimicrobial susceptibility was determined by disk diffusion (amoxicillin/clavulanic acid, cefotaxime, cefoxitine, ceftazidime, imipenem, meropenem, ciprofloxacin, gentamicin, tigecycline and colistin), and interpreted according to CLSI guidelines. The presence of blaTEM, blaSHV, blaCTX, blaKPC, blaIMP and blaVIM was screened by PCR using specific primers. Amplicons were sequenced and compared to sequences available in the GeneBank database. Results: A female patient, with 35 years old, had renal insufficiency, being an immunocompromised host. She had been submitted to ciprofloxacin and meropenem therapy...

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2014

under anaerobic conditions and it enables anaerobic growth with urocanic acid as a sole terminal ... more under anaerobic conditions and it enables anaerobic growth with urocanic acid as a sole terminal electron acceptor. The latter's capability can provide the cells of UrdA-containing bacteria with a niche where no other bacteria can compete and survive.

ABSTRACT Respiratory chains are composed of several membrane protein complexes enabling the trans... more ABSTRACT Respiratory chains are composed of several membrane protein complexes enabling the transduction of energy from oxidoreduction reactions to the establishment of transmembrene difference of the membrane potential by charge translocation. Central to aerobic respiratory chains is cytochrome bc1 complex, which presents quinol:cytochrome c oxidoreductase activity. We recognize the existence of an alternative complex , Alternative Complex III – ACIII, performing the same function as the bc 1 complex but structurally totally distinct. Besides four transmembrene proteins, ACIII contains three peripheral subunits, ActA and ActE are facing the periplasm but the orientation in relation to the membrane of the ActB remains unknown. In this work we investigated the orientation of subunit ActB and its functional implications, taking advantage of ProteinaseK activity on isolated membrane vesicles from R. marinus.

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2013

The increasing number of completely sequenced genomes led us to perform a new search for the gene... more The increasing number of completely sequenced genomes led us to perform a new search for the genes coding Available online 9 January 2013 for the different ACIII subunits. We have identified a larger number of gene clusters coding for ACIII, still confined to the bacterial domain, but extended to classes in which it was not observed before. We also found an unantic-Keywords: Alternative complex III ipated diversity in gene clusters, both in terms of its constitution and organization. The several unexpected gene CISM family arrangements brought new perspectives to the role of the different subunits of ACIII, namely in quinone binding Heme copper oxygen reductase and in proton translocation. This article is part of a Special Issue entitled: Respiratory complex III and related bc Rhodothermus marinus complexes.

Biochimica et Biophysica Acta (BBA) - Bioenergetics, 2016