Papers by Mattia Zaccarin
Introduction: Osteoarthritis (OA) is the most common joint disease. Certain neuropeptides contrib... more Introduction: Osteoarthritis (OA) is the most common joint disease. Certain neuropeptides contribute to both the continuance of symptoms and cartilage damage resulting from the disease. Symptomatic drugs are frequently used for pain control, but dissatisfaction with such an intervention has led many patients to seek other treatments, 1 of which is thermal heat from mud packs. Our intentions in the present study were to investigate a possible interaction between thermal treatment and the main neuropeptides involved in OA pathogenesis and to identify any possible detrimental effect of heat application on OA joints. Materials and Methods: Forty-eight patients with lumbar OA were enrolled in the study and randomized to Group A (12 days of mud-pack treatment at 39-40° C plus thermal bath at 37°-38° C and 500 mg acetaminophen twice per day) or Group B (drug treatment only, 500 mg acetaminophen twice per day). Blood samples were collected for assays of gamma neuropeptide (γNP), calcitonin ...
Analytical and Bioanalytical Chemistry, 2018
Surface active maghemite nanoparticles (SAMNs) are able to recognize and bind selected proteins i... more Surface active maghemite nanoparticles (SAMNs) are able to recognize and bind selected proteins in complex biological systems, forming a hard protein corona. Upon a 5-min incubation in bovine whey from mastitis-affected cows, a significant enrichment of a single peptide characterized by a molecular weight at 4338 Da originated from the proteolysis of a S1-casein was observed. Notably, among the large number of macromolecules in bovine milk, the detection of this specific peptide can hardly be accomplished by conventional analytical techniques. The selective formation of a stable binding between the peptide and SAMNs is due to the stability gained by adsorption-induced surface restructuration of the nanomaterial. We attributed the surface recognition properties of SAMNs to the chelation of iron(III) sites on their surface by sterically compatible carboxylic groups of the peptide. The specific peptide recognition by SAMNs allows its easy determination by MALDI-TOF mass spectrometry, and a threshold value of its normalized peak intensity was identified by a logistic regression approach and suggested for the rapid diagnosis of the pathology. Thus, the present report proposes the analysis of hard protein corona on nanomaterials as a perspective for developing fast analytical procedures for the diagnosis of mastitis in cows. Moreover, the huge simplification of proteome complexity by exploiting the selectivity derived by the peculiar SAMN surface topography, due to the iron(III) distribution pattern, could be of general interest, leading to competitive applications in food science and in biomedicine, allowing the rapid determination of hidden biomarkers by a cutting edge diagnostic strategy.
Archives of Biochemistry and Biophysics, 2017
Oxidation of critical signaling protein cysteines regulated by H 2 O 2 has been considered to inv... more Oxidation of critical signaling protein cysteines regulated by H 2 O 2 has been considered to involve sulfenic acid (RSOH) formation. RSOH may subsequently form either a sulfenyl amide (RSNHR') with a neighboring amide, or a mixed disulfide (RSSR') with another protein cysteine or glutathione. Previous studies have claimed that RSOH can be detected as an adduct (e.g., with 5,5-dimethylcyclohexane-1,3-dione; dimedone). Here, kinetic data are discussed which indicate that few proteins can form RSOH under physiological signaling conditions. We also present experimental evidence that indicates that (1) dimedone reacts rapidly with sulfenyl amides, and more rapidly than with sulfenic acids, and (2) that disulfides can react reversibly with amides to form sulfenyl amides. As some proteins are more stable as the sulfenyl amide than as a glutathionylated species, the former may account for some of the species previously identified as the "sulfenome"-the cellular complement of reversibly-oxidized thiol proteins generated via sulfenic acids.
Setting up of an innovative procedure for redox proteomics: and its application for definition of the redox status of cells with high metastatic potential
BACKGROUND: The cysteine (Cys) proteome includes 214.000 Cys with thiol and other forms. Of these... more BACKGROUND: The cysteine (Cys) proteome includes 214.000 Cys with thiol and other forms. Of these, only a relatively small subset functions in cell signalling. Redox-active Cys are more susceptible to oxidation, and their oxidized form is more susceptible to reduction. Specific proteomic techniques are required to identify these modifications and to study their regulation in different cell processes that are collectively known as redox proteomics. Thus, it is of interest to be able to identify both the proteins and the cysteine residues affected, and to quantify the extent of the modification involved.The quantification of differences between two or more physiological states of a biological system is among the most challenging technical tasks in proteomics: liquid chromatography coupled to mass spectrometry (LC-MS) based quantification methods have gained increasing robustness and reliability over the past five years. Many authors still share a view of redox signalling in which the fate of the cell is dependent mainly on the intensity and duration of pro-oxidant stimulus: here we sustain the involvement of an equilibrium encompassing the action of both nucleophiles and electrophiles at the same time. AIM: The dual aim of my PhD work has been both to develop suitable methodology to identify and quantify redox-active proteins in complex samples and to apply it to the study of a cellular model of breast cancer (MCF10A) engineered to reproduce malignancy. METHODS: In order to pursue this aim, I took advantage of an approach integrating differential chemical sample labelling (non-isotopic) with Cys reactive probes (NEM, IAM, HPDP) and chromatographic purification of redox-sensitive proteins, with subsequent LC-MS/MS analysis and computational data handling for OpenMS-based label-free quantification. All the steps of this methodology have been developed and validated in close collaboration with experts from both the biochemistry and bioinformatics field. RESULTS: We obtained an efficient cost-effective and isotopes-free methodology to characterize the redoxome in complex protein samples. Application of our quantification protocol to benchmark dataset leads to 100% correct estimates of under/over expression of the protein moiety. Application of the methodology to the breast cancer cellular model lead to identification of more than 300 proteins and allowed us to group-up unchanged and differentially oxidized redox-sensitive proteins in the more malignant cells in respect to their less aggressive counterpart. CONCLUSION: Despite the commonly accepted association between cancer and higher oxidative-stress, this study links higher breast cancer cells malignancy to a finely tuned dynamic equilibrium in which selected protein targets are oxidized in the context of a more reduced cell environment. Preliminary results point at the enzyme G6PDH as a crucial regulator of this redox process.
MPA: A multiple peak alignment algorithm to perform multiple comparisons of liquid-phase proteomic profiles
PROTEOMICS, 2008
Molecular and Cellular Biochemistry, 2010
Platinum-based chemotherapy, such as cisplatin, is the primary treatment for human ovarian cancer... more Platinum-based chemotherapy, such as cisplatin, is the primary treatment for human ovarian cancer. However, overcoming drug resistance has become an important issue in cancer chemotherapy. In this study, we performed 2-DE and ESI-Q-TOF MS/MS analysis to identify differential proteins expression between cisplatinsensitive (A2780S) and cisplatin-resistant (A2780-CP) ovarian cancer cell lines. Of the 14 spots identified as differentially expressed (±over twofold, P \ 0.05) between the two cell lines, ten spots (corresponding to ten unique proteins) were positively identified by ESI-Q-TOF MS/MS analysis. These proteins include capsid glycoprotein, fructose-bisphosphate aldolase C, heterogeneous nuclear ribonucleoproteins A2/B1, putative RNA-binding protein 3, Ran-specific GTPase-activating protein, ubiquitin carboxyl-terminal hydrolase isozyme L1, stathmin, ATPSH protein, chromobox protein homolog3 and phosphoglycerate kinase 1. The proteins identified in this study would be useful in revealing the mechanisms underlying cisplatin resistance and also provide some clues for further research.
Journal of Chromatography B, 2008
The effect of 100 nM sodium selenite supplementation was studied on LNCaP cells by a proteomic ap... more The effect of 100 nM sodium selenite supplementation was studied on LNCaP cells by a proteomic approach, on ProteomeLab TM PF 2D platform. Proteins were separated by liquid phase bi-dimensional chromatography and analyzed by pair-wise alignment of peaks to detect those differentially expressed. Differential expression threshold was set at a twice difference level and proteins matching this criterion were identified by MALDI-TOF and confirmed by ESI-ion trap MS/MS. Not all differentially expressed proteins found by PF 2D could be identified by MS analysis, the sensitivity of which emerging as the limiting factor. Thus, only the most abundant proteins, differently expressed following selenium supplementation, were identified. We positively showed an increase of expression of thioredoxin reductase 1, enolase 1, phosphoglycerate mutase 1, glyceraldehyde-3-phosphate dehydrogenase, heterogeneous nuclear ribonucleoprotein A2/B1, isoform A2, Ras-GTPase-activating protein SH3-domain-binding protein and Keratin 18 and a decrease of expression of peroxiredoxin 1 and heat shock protein 70, protein 8, isoform 1. Results are consistent, at least in part, with the less oxidant environment brought about by the synthesis of Se-dependent peroxidases, keeping low the steady-state concentration of hydrogen peroxide.
Free Radical Biology and Medicine, 2014
Reversible oxidation of cysteine residues is a relevant posttranslational modification of protein... more Reversible oxidation of cysteine residues is a relevant posttranslational modification of proteins. However, the low activation energy and transitory nature of the redox switch and the intrinsic complexity of the analysis render quite challenging the aim of a rigorous high-throughput screening of the redox status of redox-sensitive cysteine residues. We describe here a quantitative workflow for redox proteomics, where the ratio between the oxidized forms of proteins in the control vs treated samples is determined by a robust label-free approach. We critically present the convenience of the procedure by specifically addressing the following aspects: (i) the accurate ratio, calculated from the whole set of identified peptides rather than just isotope-tagged fragments; (ii) the application of a robust analytical pipeline to frame the most consistent data averaged over the biological variability; (iii) the relevance of using stringent criteria of analysis, even at the cost of losing potentially interesting but statistically uncertain data. The pipeline has been assessed on red blood cell membrane challenged with diamide as a model of a mild oxidative condition. The cluster of identified proteins encompassed components of the cytoskeleton more oxidized. Indirectly, our analysis confirmed the previous observation that oxidized hemoglobin binds to membranes while oxidized peroxiredoxin 2 loses affinity.
Archives of biochemistry and biophysics, Jan 13, 2016
Reversible oxidation of Cys residues is a crucial element of redox homeostasis and signaling. Acc... more Reversible oxidation of Cys residues is a crucial element of redox homeostasis and signaling. According to a popular concept in oxidative stress signaling, the oxidation of targets of signals can only take place following an overwhelming of the cellular antioxidant capacity. This concept, however, ignores the activation of feedback mechanisms possibly leading to a paradoxical effect. In a model of cancer stem cells (CSC), stably overexpressing the TAZ oncogene, we observed that the increased formation of oxidants is associated with a globally more reduced state of proteins. Redox proteomics revealed that several proteins, capable of undergoing reversible redox transitions, are indeed more reduced while just few are more oxidized. Among the proteins more oxidized, G6PDH emerges as both more expressed and activated by oxidation. This accounts for the observed more reduced state of the NADPH/NADP(+) couple. The dynamic redox flux generating this apparently paradoxical effect is rationa...
Improvements to polar 2-D electrophoresis for proteomic applications
Amino Acids, 2014
Recently, we reported a new way of performing 2-DE, called P-dimensional electrophoresis (2-PE). ... more Recently, we reported a new way of performing 2-DE, called P-dimensional electrophoresis (2-PE). In this approach, the second dimension is achieved in a radial gel which can accommodate up to six 7 cm long IPG strips simultaneously, improving reproducibility and throughput power in respect to 2-DE. Nevertheless, 2-PE was up to now limited to the use of only short strips because of technical difficulties. Here, we describe how to load longer strips (e.g., 18-24 cm) on 2-PE and report some representative images for a qualitative assessment.
Protein corona as a proteome fingerprint: The example of hidden biomarkers for cow mastitis
Colloids and Surfaces B: Biointerfaces, 2016
Proteome modifications in a biological fluid can potentially indicate the occurrence of pathologi... more Proteome modifications in a biological fluid can potentially indicate the occurrence of pathologies, even if the identification of a proteome fingerprint correlated to a specific disease represents a very difficult task. When a nanomaterial is introduced into a biological fluid, macromolecules compete to form a protein corona on the nanoparticle surface, and depending on the specific proteome, different patterns of proteins will form the final protein corona shell depending on their affinity for the nanoparticle surface. Novel surface active maghemite nanoparticles (SAMNs) display a remarkable selectivity toward protein corona formation, and they are able to concentrate proteins and peptides presenting high affinities for their surface even if they are present in very low amounts. Thus, SAMNs may confer visibility to hidden biomarkers correlated to the occurrence of a pathology. In the present report, SAMNs were introduced into milk samples from healthy cows and from animals affected by mastitis, and the selectively bound protein corona shell was easily analyzed and quantified by gel electrophoresis and characterized by mass spectrometry. Upon incubation in mastitic milk, SAMNs were able to selectively bind αs2-casein fragments containing the FALPQYLK sequence, as part of the larger casocidin-1 peptide with strong antibacterial activity, which were not present in healthy samples. Thus, SAMNs can be used as a future candidate for the rapid diagnosis of mastitis in bovine milk. The present report proposes protein competition for SAMN protein corona formation as a means of mirroring proteome modifications. Thus, the selected protein shell on the nanoparticles results in a fingerprint of the specific pathology.
Selenocysteine oxidation in glutathione peroxidase catalysis: an MS-supported quantum mechanics study
Free Radical Biology and Medicine, 2015
Glutathione peroxidases (GPxs) are enzymes working with either selenium or sulfur catalysis. They... more Glutathione peroxidases (GPxs) are enzymes working with either selenium or sulfur catalysis. They adopted diverse functions ranging from detoxification of H2O2 to redox signaling and differentiation. The relative stability of the selenoenzymes, however, remained enigmatic in view of the postulated involvement of a highly unstable selenenic acid form during catalysis. Nevertheless, density functional theory calculations obtained with a representative active site model verify the mechanistic concept of GPx catalysis and underscore its efficiency. However, they also allow that the selenenic acid, in the absence of the reducing substrate, reacts with a nitrogen in the active site. MS/MS analysis of oxidized rat GPx4 complies with the predicted structure, an 8-membered ring, in which selenium is bound as selenenylamide to the protein backbone. The intermediate can be re-integrated into the canonical GPx cycle by glutathione, whereas, under denaturing conditions, its selenium moiety undergoes β-cleavage with formation of a dehydro-alanine residue. The selenenylamide bypass prevents destruction of the redox center due to over-oxidation of the selenium or its elimination and likely allows fine-tuning of GPx activity or alternate substrate reactions for regulatory purposes.
Journal of Diabetes Research, 2015
The glucose-regulated protein94 (Grp94) has been found in complexes with IgG in plasma of Type 1 ... more The glucose-regulated protein94 (Grp94) has been found in complexes with IgG in plasma of Type 1 (T1) diabetic subjects; however, the pathogenetic meaning of Grp94-IgG complexes has not yet been elucidated. To shed light on the nature and structure of these complexes in vivo, we conducted a proteomic analysis on plasma of both T1 diabetic subjects and healthy control subjects. IgG purified from plasma was submitted to 2D PAGE followed by Western blotting and mass analysis. Grp94 was detected in plasma of all diabetic but not control subjects and found linked with its N-terminus to the IgG heavy chain. Mass analysis of heavy chain of IgG that binds Grp94 also in vitro, forming stable complexes with characteristics similar to those of native ones, permitted identifying CH2 and CH3 regions as those involved in binding Grp94. At the electron microscopy, IgG from diabetic plasma appeared as fibrils of various lengthes and dimensions, suggestive of elevated aggregating tendency conferred to IgG by Grp94. The nonimmune nature of complexes turned out to be responsible for the particular stability and structure adopted by complexes in plasma of diabetic subjects. Results are of relevance to understanding the pathogenetic mechanisms underlying diabetes and its complications.
Journal of Proteome Research, 2011
Neuroblastoma is one of the most aggressive solid tumors in the childhood. Therapy resistance to ... more Neuroblastoma is one of the most aggressive solid tumors in the childhood. Therapy resistance to anticancer drugs represents the major limitation to the effectiveness of clinical treatment. To better understand the mechanisms underlying cisplatin resistance, we performed a comparative proteomic study of the human neuroblastoma cell line SH-SY5Y and its cisplatin resistant counterpart by both the classical 2-DE electrophoresis coupled to mass spectrometry and the more innovative label-free nLC-MS E . The differentially expressed proteins were classified by bioinformatic tools according to their biological functions and their involvement in several cellular processes. Moreover, a meta-mining investigation of protein ontologies was also performed on available data from previously published proteomics studies to highlight the modulation of significant cellular pathways, which may regulate the sensitivity of neuroblastoma to cisplatin. In particular, we hypothesized a major role of the transcription factor nuclear factor-erythroid 2-related factor 2 (Nrf2) pathway. Confocal microscopy experiments, enzyme assay, and Western blotting of proteins regulated by Nrf2 provided evidences that this pathway, playing a protective role in normal cells, may represent a potential novel target to control cisplatin resistance in neuroblastoma. Cell Culture. The human neuroblastoma (NB) cell line SH-SY5Y was maintained in DMEM High glucose (GIBCO, Paisley, U.K.) containing 10% bovine serum albumin (FBS) (GIBCO), 2 mM L-glutamine (GIBCO), 1% NEAA (GIBCO), 1% sodium pyruvate (GIBCO), 10 mM HEPES (GIBCO) and 1% antibiotics (100 U/mL penicillin/streptomycin) (GIBCO) under standard conditions (37°C temperature, 5% CO 2 in a humidified atmosphere). The SH-SY5Y cell line resistant to cisplatin (SIGMA, St. Louis, MO) was selected treating cells with increasing concentration of drug starting from 1 nM to 1 µM (0.3 µg/ mL). Preparations of cell lysates for proteomic analysis were repeated in identical conditions of cell growth, 48 h after cell plating when cells reached a 80-90% confluence in 100 × 20 mm culture disks in complete media.
Improvements to polar 2-D electrophoresis for proteomic applications
Amino Acids, 2014
Recently, we reported a new way of performing 2-DE, called P-dimensional electrophoresis (2-PE). ... more Recently, we reported a new way of performing 2-DE, called P-dimensional electrophoresis (2-PE). In this approach, the second dimension is achieved in a radial gel which can accommodate up to six 7 cm long IPG strips simultaneously, improving reproducibility and throughput power in respect to 2-DE. Nevertheless, 2-PE was up to now limited to the use of only short strips because of technical difficulties. Here, we describe how to load longer strips (e.g., 18-24 cm) on 2-PE and report some representative images for a qualitative assessment.
European Journal of Cancer Supplements, 2007
Free Radical Biology and Medicine, 2015
GPx8 is a mammalian Cys--glutathione peroxidase of the endoplasmic reticulum membrane, involved i... more GPx8 is a mammalian Cys--glutathione peroxidase of the endoplasmic reticulum membrane, involved in protein folding. Its regulation is mostly unknown. We addressed both, functionality of two hypoxia response elements (HREs) within the promoter, GPx8--HRE1 and GPx8--HRE2 and the GPx8 physiological role. In HeLa cells, treatment with HIFα stabilizers, such as diethyl succinate (DES) or 2--2'-bipyridyl (BP) induces GPx8 mRNA 1.5 fold. Luciferase activity of pGL3 GPx8wt , containing a fragment of the GPx8 promoter including the two HREs, is also induced by DES/BP or by overexpressing either individual HIFα subunit. Mutating GPx8--HRE1 within pGL3 GPx8wt resulted in a significantly higher inhibition of luciferase activity than mutating GPx8--HRE2. EMSA analysis showed that both HREs exhibit enhanced binding to a nuclear extract from DES/BP--treated cells, with stronger binding by GPx8--HRE1. In DES--treated cells transfected with pGL3 GPx8wt or mutants thereof, silencing of HIF2α, but not HIF1α, abolishes luciferase activity. Thus GPx8 is a novel HIF target preferentially responding to HIF2α binding at its two novel functional GPx8--HREs, with GPx8--HRE1 playing the major role. FGF treatment increases GPx8 mRNA expression and reporter gene experiments indicate that induction occurs via HIF. Comparing the effect of depleting GPx8 on the downstream effectors of FGF or insulin signaling, revealed that absence of GPx8 results in a 16 or 12 fold increase of phosphorylated ERK 1/2 --by FGF or insulin treatment respectively. Furthermore, in GPx8 depleted cells, phosphorylation of AKT by insulin treatment increases 2.5 fold. We suggest that induction of GPx8 expression by HIF slows down proliferative signaling during hypoxia and/or growth stimulation through receptor tyrosine kinases.
Free Radical Biology and Medicine, 2015
The glutathione peroxidase homologs (GPxs) efficiently reduce hydroperoxides using electrons from... more The glutathione peroxidase homologs (GPxs) efficiently reduce hydroperoxides using electrons from glutathione (GSH), thioredoxin (Trx), or protein disulfide isomerase (PDI). Trx is preferentially used by the GPxs of the majority of bacteria, invertebrates, plants, and fungi. GSH or PDI, instead, is preferentially used by vertebrate GPxs that operate by Sec or Cys catalysis, respectively. Mammalian GPx7 and GPx8 are unique homologs that contain a peroxidatic Cys (C P ). Being reduced by PDI and located within the endoplasmic reticulum (ER), these enzymes have been involved in oxidative protein folding. Kinetic analysis indicates that oxidation of PDI by recombinant GPx7 occurs at a much faster rate than that of GSH. Nonetheless, activity measurement suggests that, at physiological concentrations, a competition between these two substrates takes place, with the rate of PDI oxidation by GPx7 controlled by the concentration of GSH, whereas the GSSG produced in the competing reaction contributes to the ER redox buffer. A mechanism has been proposed for GPx7 involving two Cys residues, in which an intramolecular disulfide of the C P is formed with an alleged resolving Cys (C R ) located in the strongly conserved FPCNQ motif (C86 in humans), a noncanonical position in GPxs. Kinetic measurements and comparison with the other thiol peroxidases containing a functional C R suggest that a resolving function of C86 in the catalytic cycle is very unlikely. We propose that GPx7 is catalytically active as a 1-Cys-GPx, in which C P both reduces H 2 O 2 and oxidizes PDI, and that the C P -C86 disulfide has instead the role of stabilizing the oxidized peroxidase in the absence of the reducing substrate.
BMC Microbiology, 2015
Background: Legumes establish with rhizobial bacteria a nitrogen-fixing symbiosis which is of the... more Background: Legumes establish with rhizobial bacteria a nitrogen-fixing symbiosis which is of the utmost importance for both plant nutrition and a sustainable agriculture. Calcium is known to act as a key intracellular messenger in the perception of symbiotic signals by both the host plant and the microbial partner. Regulation of intracellular free Ca 2+ concentration, which is a fundamental prerequisite for any Ca 2+ -based signalling system, is accomplished by complex mechanisms including Ca 2+ binding proteins acting as Ca 2+ buffers. In this work we investigated the occurrence of Ca 2+ binding proteins in Mesorhizobium loti, the specific symbiotic partner of the model legume Lotus japonicus. Results: A soluble, low molecular weight protein was found to share several biochemical features with the eukaryotic Ca 2+ -binding proteins calsequestrin and calreticulin, such as Stains-all blue staining on SDS-PAGE, an acidic isoelectric point and a Ca 2+ -dependent shift of electrophoretic mobility. The protein was purified to homogeneity by an ammonium sulfate precipitation procedure followed by anion-exchange chromatography on DEAE-Cellulose and electroendosmotic preparative electrophoresis. The Ca 2+ binding ability of the M. loti protein was demonstrated by 45 Ca 2+ -overlay assays. ESI-Q-TOF MS/MS analyses of the peptides generated after digestion with either trypsin or endoproteinase AspN identified the rhizobial protein as ferredoxin II and confirmed the presence of Ca 2+ adducts.
Journal of Chromatography B, 2008
The effect of 100 nM sodium selenite supplementation was studied on LNCaP cells by a proteomic ap... more The effect of 100 nM sodium selenite supplementation was studied on LNCaP cells by a proteomic approach, on ProteomeLab TM PF 2D platform. Proteins were separated by liquid phase bi-dimensional chromatography and analyzed by pair-wise alignment of peaks to detect those differentially expressed. Differential expression threshold was set at a twice difference level and proteins matching this criterion were identified by MALDI-TOF and confirmed by ESI-ion trap MS/MS. Not all differentially expressed proteins found by PF 2D could be identified by MS analysis, the sensitivity of which emerging as the limiting factor. Thus, only the most abundant proteins, differently expressed following selenium supplementation, were identified. We positively showed an increase of expression of thioredoxin reductase 1, enolase 1, phosphoglycerate mutase 1, glyceraldehyde-3-phosphate dehydrogenase, heterogeneous nuclear ribonucleoprotein A2/B1, isoform A2, Ras-GTPase-activating protein SH3-domain-binding protein and Keratin 18 and a decrease of expression of peroxiredoxin 1 and heat shock protein 70, protein 8, isoform 1. Results are consistent, at least in part, with the less oxidant environment brought about by the synthesis of Se-dependent peroxidases, keeping low the steady-state concentration of hydrogen peroxide.
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Papers by Mattia Zaccarin