Publications by authors named "Andrea Scaloni"

265 Publications

Carbonic Anhydrases: New Perspectives on Protein Functional Role and Inhibition in .

Front Microbiol 2021 19;12:629163. Epub 2021 Mar 19.

Department of Biology, Agriculture and Food Sciences, National Research Council (CNR), Institute of Biosciences and Bioresources, Naples, Italy.

Our understanding of the function of bacterial carbonic anhydrases (CAs, EC 4.2.1.1) has increased significantly in the last years. CAs are metalloenzymes able to modulate CO, HCO and H concentration through their crucial role in catalysis of reversible CO hydration (CO + HO ⇄ HCO + H). In all living organisms, CA activity is linked to physiological processes, such as those related to the transport and supply of CO or HCO, pH homeostasis, secretion of electrolytes, biosynthetic processes and photosynthesis. These important processes cannot be ensured by the very low rate of the non-catalyzed reaction of CO hydration. It has been recently shown that CAs are important biomolecules for many bacteria involved in human infections, such as , , , , and . In these species, CA activity promotes microorganism growth and adaptation in the host, or modulates bacterial toxin production and virulence. In this review, recent literature in this research field and some of the above-mentioned issues are discussed, namely: () the implication of CAs from bacterial pathogens in determining the microorganism growth and virulence; () the druggability of these enzymes using classical CA inhibitors (CAIs) of the sulfonamide-type as examples; () the role played by CAs in the acid tolerance/adaptation of the microbe within the human abdomen; () the role of CAs played in the outer membrane vesicles spawned by in its planktonic and biofilm phenotypes; () the possibility of using CAIs in combination with probiotic strains as a novel anti-ulcer treatment approach. The latter approach may represent an innovative and successful strategy to fight gastric infections in the era of increasing resistance of pathogenic bacteria to classical antibiotics.
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http://dx.doi.org/10.3389/fmicb.2021.629163DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8017301PMC
March 2021

Adaptive Thermogenesis Driving Catch-Up Fat Is Associated With Increased Muscle Type 3 and Decreased Hepatic Type 1 Iodothyronine Deiodinase Activities: A Functional and Proteomic Study.

Front Endocrinol (Lausanne) 2021 4;12:631176. Epub 2021 Mar 4.

Department of Science and Technologies, University of Sannio, Benevento, Italy.

Refeeding after caloric restriction induces weight regain and a disproportionate recovering of fat mass rather than lean mass (catch-up fat) that, in humans, associates with higher risks to develop chronic dysmetabolism. Studies in a well-established rat model of semistarvation-refeeding have reported that catch-up fat associates with hyperinsulinemia, glucose redistribution from skeletal muscle to white adipose tissue and suppressed adaptive thermogenesis sustaining a high efficiency for fat deposition. The skeletal muscle of catch-up fat animals exhibits reduced insulin-stimulated glucose utilization, mitochondrial dysfunction, delayed contraction-relaxation kinetics, increased proportion of slow fibers and altered local thyroid hormone metabolism, with suggestions of a role for iodothyronine deiodinases. To obtain novel insights into the skeletal muscle response during catch-up fat in this rat model, the functional proteomes of tibialis anterior and soleus muscles, harvested after 2 weeks of caloric restriction and 1 week of refeeding, were studied. Furthermore, to assess the implication of thyroid hormone metabolism in catch-up fat, circulatory thyroid hormones as well as liver type 1 (D1) and liver and skeletal muscle type 3 (D3) iodothyronine deiodinase activities were evaluated. The proteomic profiling of both skeletal muscles indicated catch-up fat-induced alterations, reflecting metabolic and contractile adjustments in soleus muscle and changes in glucose utilization and oxidative stress in tibialis anterior muscle. In response to caloric restriction, D3 activity increased in both liver and skeletal muscle, and persisted only in skeletal muscle upon refeeding. In parallel, liver D1 activity decreased during caloric restriction, and persisted during catch-up fat at a time-point when circulating levels of T4, T3 and rT3 were all restored to those of controls. Thus, during catch-up fat, a local hypothyroidism may occur in liver and skeletal muscle despite systemic euthyroidism. The resulting reduced tissue thyroid hormone bioavailability, likely D1- and D3-dependent in liver and skeletal muscle, respectively, may be part of the adaptive thermogenesis sustaining catch-up fat. These results open new perspectives in understanding the metabolic processes associated with the high efficiency of body fat recovery after caloric restriction, revealing new implications for iodothyronine deiodinases as putative biological brakes contributing in suppressed thermogenesis driving catch-up fat during weight regain.
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http://dx.doi.org/10.3389/fendo.2021.631176DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7971177PMC
March 2021

Evidences for a Nutritional Role of Iodine in Plants.

Front Plant Sci 2021 17;12:616868. Epub 2021 Feb 17.

Plant Lab, Institute of Life Sciences, Scuola Superiore Sant'Anna, Pisa, Italy.

Little is known about the role of iodine in plant physiology. We evaluated the impact of low concentrations of iodine on the phenotype, transcriptome and proteome of . Our experiments showed that removal of iodine from the nutrition solution compromises plant growth, and restoring it in micromolar concentrations is beneficial for biomass accumulation and leads to early flowering. In addition, iodine treatments specifically regulate the expression of several genes, mostly involved in the plant defence response, suggesting that iodine may protect against both biotic and abiotic stress. Finally, we demonstrated iodine organification in proteins. Our bioinformatic analysis of proteomic data revealed that iodinated proteins identified in the shoots are mainly associated with the chloroplast and are functionally involved in photosynthetic processes, whereas those in the roots mostly belong and/or are related to the action of various peroxidases. These results suggest the functional involvement of iodine in plant nutrition.
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http://dx.doi.org/10.3389/fpls.2021.616868DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7925997PMC
February 2021

Monitoring aging of hen egg by integrated quantitative peptidomic procedures.

Food Res Int 2021 Feb 15;140:110010. Epub 2020 Dec 15.

Proteomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, Naples 80147, Italy. Electronic address:

Environmental conditions and timing of egg storage highly affect raw material quality. Aging and endogenous processing of constituent proteins can determine important changes in specific functions and technological properties of inner egg compartments. We here used integrated peptidomic procedures to identify peptide markers of egg freshness. At first, peptides extracted from egg white and yolk plasma taken from eggs stored for different times were subjected to a label-free untargeted quantitation procedure based on nanoLC-ESI-Q-Orbitrap-MS/MS, which identified 836 and 1974 unique variable molecules, respectively. By applying stringent criteria for filtering data, 30 and 66 putative egg aging markers were selected for egg white and yolk plasma, respectively. Proposed molecules were then validated through a targeted label-free parallel reaction monitoring procedure based on nanoLC-ESI-Q-Orbitrap-MS/MS, confirming quantitative trends for 19 and 25 peptides in egg white and yolk plasma, respectively, and generating a robust panel of egg storage markers. Quantitative results reflected physico-chemical phenomena occurring in egg compartments during storage and offered essential information for the development of novel control procedures to assess quality features of fresh/stored raw material.
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http://dx.doi.org/10.1016/j.foodres.2020.110010DOI Listing
February 2021

Defense against the Pathogenic Fungus Is Dependent on the Integrity of the Unfolded Protein Response.

Biomolecules 2021 Feb 8;11(2). Epub 2021 Feb 8.

Department of Biology, University of Rome Tor Vergata, 00133 Rome, Italy.

Heald & Wolf is a worldwide-spread necrotrophic fungus closely related to the genus, well-known because many member species provoke severe diseases in cereal crops and studied because they produce sesterpenoid phytoxins named ophiobolins which possess interesting biological properties. The unfolded protein response (UPR) is a conserved mechanism protecting eukaryotic cells from the accumulation of unfolded/misfolded proteins in the endoplasmic reticulum (ER). In plants, consolidated evidence supports the role of UPR in the tolerance to abiotic stress, whereas much less information is available concerning the induction of ER stress by pathogen infection and consequent UPR elicitation as part of the defense response. In this study, the infection process of in wild type and UPR-defective double mutant plants was comparatively investigated, with the aim to address the role of UPR in the expression of resistance to the fungal pathogen. The results of confocal microscopy, as well as of qRT-PCR transcript level analysis of UPR genes, proteomics, microRNAs expression profile and HPLC-based hormone analyses demonstrated that ophiobolin produced by the fungus during infection compromised ER integrity and that impairment of the IRE1/bZIP60 pathway of UPR hampered the full expression of resistance, thereby enhancing plant susceptibility to the pathogen.
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http://dx.doi.org/10.3390/biom11020240DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7915340PMC
February 2021

Identification of RNA-binding proteins that partner with Lin28a to regulate Dnmt3a expression.

Sci Rep 2021 Jan 27;11(1):2345. Epub 2021 Jan 27.

Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 80131, Naples, Italy.

Lin28 is an evolutionary conserved RNA-binding protein that plays important roles during embryonic development and tumorigenesis. It regulates gene expression through two different post-transcriptional mechanisms. The first one is based on the regulation of miRNA biogenesis, in particular that of the let-7 family, whose expression is suppressed by Lin28. Thus, loss of Lin28 leads to the upregulation of mRNAs that are targets of let-7 species. The second mechanism is based on the direct interaction of Lin28 with a large number of mRNAs, which results in the regulation of their translation. This second mechanism remains poorly understood. To address this issue, we purified high molecular weight complexes containing Lin28a in mouse embryonic stem cells (ESCs). Numerous proteins, co-purified with Lin28a, were identified by proteomic procedures and tested for their possible role in Lin28a-dependent regulation of the mRNA encoding DNA methyltransferase 3a (Dnmt3a). The results show that Lin28a activity is dependent on many proteins, including three helicases and four RNA-binding proteins. The suppression of four of these proteins, namely Ddx3x, Hnrnph1, Hnrnpu or Syncrip, interferes with the binding of Lin28a to the Dnmt3a mRNA, thus suggesting that they are part of an oligomeric ribonucleoprotein complex that is necessary for Lin28a activity.
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http://dx.doi.org/10.1038/s41598-021-81429-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7841167PMC
January 2021

Protein kinase Cα regulates the nucleocytoplasmic shuttling of KRIT1.

J Cell Sci 2021 Feb 4;134(3). Epub 2021 Feb 4.

Department of Clinical and Biological Sciences, University of Torino, 10043 Orbassano, Torino, Italy

KRIT1 is a scaffolding protein that regulates multiple molecular mechanisms, including cell-cell and cell-matrix adhesion, and redox homeostasis and signaling. However, rather little is known about how KRIT1 is itself regulated. KRIT1 is found in both the cytoplasm and the nucleus, yet the upstream signaling proteins and mechanisms that regulate KRIT1 nucleocytoplasmic shuttling are not well understood. Here, we identify a key role for protein kinase C (PKC) in this process. In particular, we found that PKC activation promotes the redox-dependent cytoplasmic localization of KRIT1, whereas inhibition of PKC or treatment with the antioxidant N-acetylcysteine leads to KRIT1 nuclear accumulation. Moreover, we demonstrated that the N-terminal region of KRIT1 is crucial for the ability of PKC to regulate KRIT1 nucleocytoplasmic shuttling, and may be a target for PKC-dependent regulatory phosphorylation events. Finally, we found that silencing of PKCα, but not PKCδ, inhibits phorbol 12-myristate 13-acetate (PMA)-induced cytoplasmic enrichment of KRIT1, suggesting a major role for PKCα in regulating KRIT1 nucleocytoplasmic shuttling. Overall, our findings identify PKCα as a novel regulator of KRIT1 subcellular compartmentalization, thus shedding new light on the physiopathological functions of this protein.
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http://dx.doi.org/10.1242/jcs.250217DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7875496PMC
February 2021

Effect of Sulfonamides and Their Structurally Related Derivatives on the Activity of ι-Carbonic Anhydrase from

Int J Mol Sci 2021 01 8;22(2). Epub 2021 Jan 8.

Institute of Biosciences and Bioresources, CNR, via Pietro Castellino 111, 80131 Napoli, Italy.

Carbonic anhydrases (CAs) are essential metalloenzymes in nature, catalyzing the carbon dioxide reversible hydration into bicarbonate and proton. In humans, breathing and many other critical physiological processes depend on this enzymatic activity. The CA superfamily function and inhibition in pathogenic bacteria has recently been the object of significant advances, being demonstrated to affect microbial survival/virulence. Targeting bacterial CAs may thus be a valid alternative to expand the pharmacological arsenal against the emergence of widespread antibiotic resistance. Here, we report an extensive study on the inhibition profile of the recently discovered ι-CA class present in some bacteria, including , namely BteCAι, using substituted benzene-sulfonamides and clinically licensed sulfonamide-, sulfamate- and sulfamide-type drugs. The BteCAι inhibition profile showed: (i) several benzene-sulfonamides with an inhibition constant lower than 100 nM; (ii) a different behavior with respect to other α, β and γ-CAs; (iii) clinically used drugs having a micromolar affinity. This prototype study contributes to the initial recognition of compounds which efficiently and selectively inhibit a bacterial member of the ι-CA class, for which such a selective inhibition with respect to other protein isoforms present in the host is highly desired and may contribute to the development of novel antimicrobials.
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http://dx.doi.org/10.3390/ijms22020571DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7827628PMC
January 2021

Anion inhibition studies of the Zn(II)-bound ι-carbonic anhydrase from the Gram-negative bacterium .

J Enzyme Inhib Med Chem 2021 Dec;36(1):372-376

Department of NEUROFARBA, Section of Pharmaceutical and Nutraceutical Sciences, University of Florence, Polo Scientifico, Firenze, Italy.

, a Gram-negative bacterium, encodes for the ι-class carbonic anhydrase (CA, EC 4.2.1.1) BteCAι, which was recently characterised. It acts as a good catalyst for the hydration of CO to bicarbonate and protons, with a k value of 3.0 × 10 s and k/K value of 3.9 × 10 M s. No inhibition data on this new class of enzymes are available to date. We report here an anion and small molecules inhibition study of BteCAι, which we prove to be a zinc(II)- and not manganese(II)-containing enzyme, as reported for diatom ι-CAs. The best inhibitors were sulphamic acid, stannate, phenylarsonic acid, phenylboronic acid and sulfamide (K values of 6.2-94 µM), whereas diethyldithiocarbamate, tellurate, selenate, bicarbonate and cyanate were submillimolar inhibitors (K values of 0.71-0.94 mM). The halides (except iodide), thiocyanate, nitrite, nitrate, carbonate, bisulphite, sulphate, hydrogensulfide, peroxydisulfate, selenocyanate, fluorosulfonate and trithiocarbonate showed K values in the range of 3.1-9.3 mM.
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http://dx.doi.org/10.1080/14756366.2020.1867122DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7782983PMC
December 2021

CA IX Stabilizes Intracellular pH to Maintain Metabolic Reprogramming and Proliferation in Hypoxia.

Front Oncol 2020 2;10:1462. Epub 2020 Sep 2.

Department of Tumor Biology, Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Bratislava, Slovakia.

Tumor hypoxia represents a severe microenvironmental stress that is frequently associated with acidosis. Cancer cells respond to these stresses with changes in gene expression that promote survival at least in part through pH regulation and metabolic reprogramming. Hypoxia-induced carbonic anhydrase IX (CA IX) plays a critical adaptive role in response to hypoxic and acidic environments by catalytically hydrating extracellular CO to produce bicarbonate for buffering intracellular pH (pHi). We used proteome-wide profiling to study the cellular response to transient CA IX knockdown in hypoxia and found a decrease in the levels of key glycolytic enzymes and lactate dehydrogenase A (LDHA). Interestingly, the activity of LDH was also decreased as demonstrated by native in-gel activity assay. These changes led to a significant reduction in glycolytic flux and extracellular lactate levels in cancer cells , contributing to a decrease in proliferation. Interestingly, addition of the alternative LDH substrate alpha-ketobutyrate restored LDHA activity, extracellular acidification, pHi, and cellular proliferation. These results indicate that in the absence of CA IX, reduction of pHi disrupts LDHA activity and hinders the cellular capacity to regenerate NAD and secrete protons to the extracellular space. Hypoxia-induced CA IX therefore mediates adaptation to microenvironmental hypoxia and acidosis directly, by enzymatically converting extracellular CO to bicarbonate, and indirectly, by maintaining glycolysis-permissive intracellular milieu.
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http://dx.doi.org/10.3389/fonc.2020.01462DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7493625PMC
September 2020

Biochar Administration to San Marzano Tomato Plants Cultivated Under Low-Input Farming Increases Growth, Fruit Yield, and Affects Gene Expression.

Front Plant Sci 2020 27;11:1281. Epub 2020 Aug 27.

Department of Science and Technology, University of Sannio, Benevento, Italy.

Biochar is a rich-carbon charcoal obtained by pyrolysis of biomasses, which was used since antiquity as soil amendant. Its storage in soils was demonstrated contributing to abate the effects of climate changes by sequestering carbon, also providing bioenergy, and improving soil characteristics and crop yields. Despite interest in this amendant, there is still poor information on its effects on soil fertility and plant growth. Considerable variation in the plant response has been reported, depending on biomass source, pyrolysis conditions, crop species, and cultivation practices. Due to these conflicting evidences, this work was aimed at studying the effects of biochar from pyrolyzed wood at 550°C, containing 81.1% carbon and 0.91% nitrogen, on growth and yield of tomato plants experiencing low-input farming conditions. San Marzano ecotype from Southern Italy was investigated, due to its renowned quality and adaptability to sustainable farming practices. Biochar administration improved vegetative growth and berry yield, while affecting gene expression and protein repertoire in berries. Different enzymes of carbon metabolism and photosynthesis were over-represented, whereas various stress-responsive and defense proteins were down-represented. Molecular results are here discussed in relation to estimated agronomic parameters to provide a rationale justifying the growth-promoting effect of this soil amendant.
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http://dx.doi.org/10.3389/fpls.2020.01281DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7481538PMC
August 2020

Absence of uncoupling protein 3 at thermoneutrality influences brown adipose tissue mitochondrial functionality in mice.

FASEB J 2020 11 18;34(11):15146-15163. Epub 2020 Sep 18.

Department of Biology, University of Naples Federico II, Naples, Italy.

The physiological role played by uncoupling protein 3 (UCP3) in brown adipose tissue (BAT) has not been fully elucidated so far. In the present study, we evaluated the impact of the absence of UCP3 on BAT mitochondrial functionality and morphology. To this purpose, wild type (WT) and UCP3 Knockout (KO) female mice were housed at thermoneutrality (30°C), a condition in which BAT contributes to energy homeostasis independently of its cold-induced thermogenic function. BAT mitochondria from UCP3 KO mice presented a lower ability to oxidize the fatty acids and glycerol-3-phosphate, and an enhanced oxidative stress as revealed by enhanced mitochondrial electron leak, lipid hydroperoxide levels, and induction of antioxidant mitochondrial enzymatic capacity. The absence of UCP3 also influenced the mitochondrial super-molecular protein aggregation, an important feature for fatty acid oxidation rate as well as for adequate cristae organization and mitochondrial shape. Indeed, electron microscopy revealed alterations in mitochondrial morphology in brown adipocytes from KO mice. In the whole, data here reported show that the absence of UCP3 results in a significant alteration of BAT mitochondrial physiology and morphology. These observations could also help to clarify some aspects of the association between metabolic disorders associated with low UCP3 levels, as previously reported in human studies.
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http://dx.doi.org/10.1096/fj.202000995RDOI Listing
November 2020

Low-protein/high-carbohydrate diet induces AMPK-dependent canonical and non-canonical thermogenesis in subcutaneous adipose tissue.

Redox Biol 2020 09 9;36:101633. Epub 2020 Jul 9.

Department Biology, University of Rome Tor Vergata, Via Della Ricerca Scientifica, Rome, Italy; IRCCS Fondazione Santa Lucia, Rome, Italy. Electronic address:

Low-protein/high-carbohydrate (LPHC) diet has been suggested to promote metabolic health and longevity in adult humans and animal models. However, the complex molecular underpinnings of how LPHC diet leads to metabolic benefits remain elusive. Through a multi-layered approach, here we observed that LPHC diet promotes an energy-dissipating response consisting in the parallel recruitment of canonical and non-canonical (muscular) thermogenic systems in subcutaneous white adipose tissue (sWAT). In particular, we measured Ucp1 induction in association with up-regulation of actomyosin components and several Serca (Serca1, Serca2a, Serca2b) ATPases. In beige adipocytes, we observed that AMPK activation is responsible for transducing the amino acid lowering in an enhanced fat catabolism, which sustains both Ucp1-and Serca-dependent energy dissipation. Limiting AMPK activation counteracts the expression of brown fat and muscular genes, including Ucp1 and Serca, as well as mitochondrial oxidative genes. We observed that mitochondrial reactive oxygen species are the upstream molecules controlling AMPK-mediated metabolic rewiring in amino acid-restricted beige adipocytes. Our findings delineate a novel metabolic phenotype of responses to amino acid shortage, which recapitulates some of the benefits of cool temperature in sWAT. In conclusion, this highlights LPHC diet as a valuable and practicable strategy to prevent metabolic diseases through the enhancement of mitochondrial oxidative metabolism and the recruitment of different energy dissipating routes in beige adipocytes.
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http://dx.doi.org/10.1016/j.redox.2020.101633DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7358542PMC
September 2020

Analysis of post-translational modifications in soluble proteins involved in chemical communication from mammals and insects.

Authors:
Andrea Scaloni

Methods Enzymol 2020 18;642:103-124. Epub 2020 May 18.

Proteomics and Mass Spectrometry Laboratory, ISPAAM, National Research Council, Naples, Italy. Electronic address:

Different soluble, small polypeptide molecules isolated from biological tissues/body fluids of mammals and insects have been reported to bind odorant compounds and pheromones, and have been accordingly named odorant-binding proteins, pheromone-binding proteins and chemosensory proteins. Mammal and insect odorant-binding and pheromone-binding proteins show evident differences in their primary structure; in insects, the same condition occurs for odorant- and pheromone-binding proteins, which belong to the same protein family, and chemosensory proteins. In order to understand their molecular characteristics, these proteins have been subjected to an extensive analysis for their post-translational modifications through mass spectrometry approaches. Depending on proteins, they have been characterized as having specific disulfide bridge arrangements, which were consistent among molecular sequence homologs. Only selected odorant-binding proteins from mammals have been demonstrated bearing N-linked glycan structures and other modifications. In this review article, we discuss the application of mass spectrometry procedures for the analysis of post-translational modifications in odorant-binding proteins, pheromone-binding proteins and chemosensory proteins. Different example proteins are reported, and protocols are presented for obtaining definitive information in this context.
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http://dx.doi.org/10.1016/bs.mie.2020.04.062DOI Listing
May 2020

Novel Biomarkers of Mastitis in Goat Milk Revealed by MALDI-TOF-MS-Based Peptide Profiling.

Biology (Basel) 2020 Jul 28;9(8). Epub 2020 Jul 28.

Institute for the Animal Production System in the Mediterranean Environment (ISPAAM), National Research Council (CNR), 80147 Naples, Italy.

Mastitis is the most common infection of dairy goats impairing milk production and quality, which is usually recognized by mammary gland visual inspection and palpation. Subclinical forms of the disease are also widely represented, which lack the typical signs of the clinical ones but are still associated with reduced production and safety for human consumption of milk, generally presenting a high bacterial count. In order to obtain novel analytical tools for rapid and non-invasive diagnosis of mastitis in goats, we analyzed milk samples from healthy, subclinical and clinical mastitic animals with a MALDI-TOF-MS-based peptidomic platform, generating disease group-specific spectral profiles whose signal intensity and mass values were analyzed by statistics. Peculiar spectral signatures of mastitis with respect to the control were identified, while no significant spectral differences were observed between clinical and subclinical milk samples. Discriminant signals were assigned to specific peptides through nanoLC-ESI-Q-Orbitrap-MS/MS experiments. Some of these molecules were predicted to have an antimicrobial activity based on their strong similarity with homolog bioactive compounds from other mammals. Through the definition of a panel of peptide biomarkers, this study provides a very rapid and low-cost method to routinely detect mastitic milk samples even though no evident clinical signs in the mammary gland are observed.
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http://dx.doi.org/10.3390/biology9080193DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7464427PMC
July 2020

Applications on Strawberry Plants Modulate the Physiological Processes Positively Affecting Fruit Production and Quality.

Front Microbiol 2020 3;11:1364. Epub 2020 Jul 3.

Institute for Sustainable Plant Protection, National Research Council, Portici, Italy.

Many spp. are successful plant beneficial microbial inoculants due to their ability to act as biocontrol agents with direct antagonistic activities to phytopathogens, and as biostimulants capable of promoting plant growth. This work investigated the effects of treatments with three selected strains (T22, TH1, and GV41) to strawberry plants on the productivity, metabolites and proteome of the formed fruits. applications stimulated plant growth, increased strawberry fruit yield, and favored selective accumulation of anthocyanins and other antioxidants in red ripened fruits. Proteomic analysis of fruits harvested from the plants previously treated with demonstrated that the microbial inoculants highly affected the representation of proteins associated with responses to stress/external stimuli, nutrient uptake, protein metabolism, carbon/energy metabolism and secondary metabolism, also providing a possible explanation to the presence of specific metabolites in fruits. Bioinformatic analysis of these differential proteins revealed a central network of interacting molecular species, providing a rationale to the concomitant modulation of different plant physiological processes following the microbial inoculation. These findings indicated that the application of -based products exerts a positive impact on strawberry, integrating well with previous observations on the molecular mechanisms activated in roots and leaves of other tested plant species, demonstrating that the efficacy of using a biological approach with beneficial microbes on the maturing plant is also able to transfer advantages to the developing fruits.
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http://dx.doi.org/10.3389/fmicb.2020.01364DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7350708PMC
July 2020

OctoPartenopin: Identification and Preliminary Characterization of a Novel Antimicrobial Peptide from the Suckers of .

Mar Drugs 2020 Jul 23;18(8). Epub 2020 Jul 23.

Department of Pharmacy, University of Naples "Federico II", Via Mezzocannone 16, 80134 Naples, Italy.

Microorganism resistance to conventional antibiotics represents one of the major global health concerns. This paper focuses on a peptide (OctoPartenopin) extracted from suckers of bioassay-guided chromatographic fractionation was used to identify this sequence, which holds significant antibacterial activity against Gram-positive and Gram-negative bacteria. OctoPartenopin is encrypted within the calponin sequence and was associated with the high levels of proteolytic activity already reported in octopus arm suckers. We synthesized the parent peptide and four analogues; all peptide were tested for their antibacterial and antibiofilm activities. Preliminary antibiofilm experiments showed that that one of the analogues had the best activity in both inhibition and eradication of biofilm of all three microorganisms tested. The occurrence of OctoPartenopin in arm suckers provided novel speculative information on animal behavior, as concerns maternal care of fertilized eggs. Our results highlight that suckers are a rich source of multifaceted peptides to develop alternative antimicrobial agents and food preservatives.
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http://dx.doi.org/10.3390/md18080380DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7460285PMC
July 2020

Proteolysis and Process-Induced Modifications in Synbiotic Yogurt Investigated by Peptidomics and Phosphopeptidomics.

J Agric Food Chem 2020 Aug 31;68(32):8744-8754. Epub 2020 Jul 31.

Proteomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147 Naples, Italy.

Probiotic and synbiotic yogurt preparations were manufactured at the semi-industrial pilot scale with and strains without inulin or fortified with 1 and 3% (w/w) inulin. The pathway of casein breakdown was determined in probiotic, synbiotic, conventional yogurt, and nonstarted milk base using HPLC-ESI-MS/MS-based peptidomics and phosphopeptidomics; in the latter case, casein phosphorylated peptides (CPPs) were previously enriched by hydroxyapatite chromatography. Compared with traditional yogurt, casein proteolysis increased in probiotic and even more in synbiotic yogurt with 1% inulin. Fortification with 3% inulin greatly modified the proteolytic pattern, indicating a characteristic contribution of probiotics to proteolysis. The enhanced proteolysis in synbiotic yogurt exposed the neo-formed peptides to progressively increase enzymatic or chemical modifications, such as dephosphorylation of CPPs, methionine oxidation, and formation of N-terminal pyroglutamic acids. These modifications might constitute molecular signature descriptors of metabolic processes mediated by complex bacterial communities, with technological, nutritional, and sensorial significance.
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http://dx.doi.org/10.1021/acs.jafc.0c02603DOI Listing
August 2020

Human carbonic anhydrases and post-translational modifications: a hidden world possibly affecting protein properties and functions.

J Enzyme Inhib Med Chem 2020 Dec;35(1):1450-1461

Istituto di Biostrutture e Bioimmagini-National Research Council, Napoli, Italy.

Human carbonic anhydrases (CAs) have become a well-recognized target for the design of inhibitors and activators with biomedical applications. Accordingly, an enormous amount of literature is available on their biochemical, functional and structural aspects. Nevertheless post-translational modifications (PTMs) occurring on these enzymes and their functional implications have been poorly investigated so far. To fill this gap, in this review we have analysed all PTMs occurring on human CAs, as deriving from the search in dedicated databases, showing a widespread occurrence of modification events in this enzyme family. By combining these data with sequence alignments, inspection of 3 D structures and available literature, we have summarised the possible functional implications of these PTMs. Although in some cases a clear correlation between a specific PTM and the CA function has been highlighted, many modification events still deserve further dedicated studies.
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http://dx.doi.org/10.1080/14756366.2020.1781846DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7470082PMC
December 2020

Anion Inhibition Studies of the Beta-Carbonic Anhydrase from .

Molecules 2020 May 31;25(11). Epub 2020 May 31.

Institute of Biosciences and Bioresources, CNR, Via Pietro Castellino 111, 80131 Napoli, Italy.

The interconversion of CO and HCO is catalyzed by a superfamily of metalloenzymes, known as carbonic anhydrases (CAs, EC 4.2.1.1), which maintain the equilibrium between dissolved inorganic CO and HCO. In the genome of , a Gram-negative bacterium typically colonizing the lower intestine of warm-blooded organisms, the cyn operon gene includes the CynT gene, encoding for a β-CA, and CynS gene, encoding for the cyanase. CynT (β-CA) prevents the depletion of the cellular bicarbonate, which is further used in the reaction catalyzed by cyanase. A second β-CA (CynT2 or Can or yadF), as well as a γ and ι-CAs were also identified in the genome. CynT2 is essential for bacterial growth at atmospheric CO concentration. Here, we characterized the kinetic properties and the anion inhibition profiles of recombinant CynT2. The enzyme showed a good activity for the physiological CO hydratase reaction with the following parameters: k = 5.3 × 10 s and k/K = of 4.1 × 10 M s. Sulfamide, sulfamate, phenylboronic acid, phenylarsonic acid, and diethyldithiocarbamate were the most effective CynT2 inhibitors (K = 2.5 to 84 µM). The anions allowed for a detailed understanding of the interaction of inhibitors with the amino acid residues surrounding the catalytic pocket of the enzyme and may be used as leads for the design of more efficient and specific inhibitors.
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http://dx.doi.org/10.3390/molecules25112564DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7321114PMC
May 2020

Influence of the Casein Composite Genotype on Milk Quality and Coagulation Properties in the Endangered Agerolese Cattle Breed.

Animals (Basel) 2020 May 20;10(5). Epub 2020 May 20.

Department of Veterinary Medicine and Animal Production, University of Naples Federico II, via Delpino 1, 80137 Naples, Italy.

The aim of this study was the characterization of CSN1S1, CSN2 and CSN3 genetic variability in Agerolese cattle, and the investigation of the effect of casein composite genotypes (CSN1S1, CSN2 and CSN3) on quality and coagulation traits of the corresponding milk. To these purposes, blood and milk from 84 cows were sampled and analysed. Allele frequencies at and revealed no Hardy-Weinberg equilibrium in the population with a prevalence of allele A for and allele B for . BBAAAB and BBAAAB composite genotypes were the most common in the population. BBAAAB showed a higher total solids and fat content (12.70 ± 0.16 and 3.93 ± 0.10, respectively), while BBAABB showed the best coagulation properties (RCT 12.62 ± 0.81; k 5.84 ± 0.37; a 23.72 ± 1.10). Interestingly, the A allele of was very widespread in the population; thus, it will be intriguing to verify if AA Agerolese cattle milk and the derived cheese may have better nutraceutical characteristics.
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http://dx.doi.org/10.3390/ani10050892DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7278449PMC
May 2020

Effect of Bioactive Metabolite Treatments on the Production, Quality, and Protein Profile of Strawberry Fruits.

J Agric Food Chem 2020 Jul 24;68(27):7246-7258. Epub 2020 Jun 24.

Institute for Sustainable Plant Protection, National Research Council, 80055 Portici, Naples, Italy.

Fungi of the genus produce secondary metabolites having several biological activities that affect plant metabolism. We examined the effect of three bioactive metabolites (BAMs), namely, 6-pentyl-α-pyrone (6PP), harzianic acid (HA), and hydrophobin 1 (HYTLO1), on yield, fruit quality, and protein representation of strawberry plants. In particular, 6PP and HA increased the plant yield and number of fruits, when compared to control, while HYTLO1 promoted the growth of the roots and increased the total soluble solids content up to 19% and the accumulation of ascorbic acid and cyanidin 3--glucoside in red ripened fruits. Proteomic analysis showed that BAMs influenced the representation of proteins associated with the protein metabolism, response to stress/external stimuli, vesicle trafficking, carbon/energy, and secondary metabolism. Results suggest that the application of BAMs affects strawberry plant productivity and fruit quality and integrate previous observations on deregulated molecular processes in roots and leaves of -treated plants with original data on fruits.
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http://dx.doi.org/10.1021/acs.jafc.0c01438DOI Listing
July 2020

Cleavage of the APE1 N-Terminal Domain in Acute Myeloid Leukemia Cells Is Associated with Proteasomal Activity.

Biomolecules 2020 03 31;10(4). Epub 2020 Mar 31.

Department of Medicine, University of Udine, 33100 Udine, Italy.

Apurinic/apyrimidinic endonuclease 1 (APE1), the main mammalian AP-endonuclease for the resolution of DNA damages through the base excision repair (BER) pathway, acts as a multifunctional protein in different key cellular processes. The signals to ensure temporo-spatial regulation of APE1 towards a specific function are still a matter of debate. Several studies have suggested that post-translational modifications (PTMs) act as dynamic molecular mechanisms for controlling APE1 functionality. Interestingly, the N-terminal region of APE1 is a disordered portion functioning as an interface for protein binding, as an acceptor site for PTMs and as a target of proteolytic cleavage. We previously demonstrated a cytoplasmic accumulation of truncated APE1 in acute myeloid leukemia (AML) cells in association with a mutated form of nucleophosmin having aberrant cytoplasmic localization (NPM1c+). Here, we mapped the proteolytic sites of APE1 in AML cells at Lys31 and Lys32 and showed that substitution of Lys27, 31, 32 and 35 with alanine impairs proteolysis. We found that the loss of the APE1 N-terminal domain in AML cells is dependent on the proteasome, but not on granzyme A/K as described previously. The present work identified the proteasome as a contributing machinery involved in APE1 cleavage in AML cells, suggesting that acetylation can modulate this process.
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http://dx.doi.org/10.3390/biom10040531DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7226146PMC
March 2020

The Small ORF Stimulates Growth and Morphological Development and Exerts Opposite Effects on Actinorhodin and Calcium-Dependent Antibiotic Production.

Front Microbiol 2020 19;11:224. Epub 2020 Feb 19.

Laboratory of Molecular Microbiology and Biotechnology, STEBICEF Department, University of Palermo, Palermo, Italy.

In actinomycetes, antibiotic production is often associated with a morpho-physiological differentiation program that is regulated by complex molecular and metabolic networks. Many aspects of these regulatory circuits have been already elucidated and many others still deserve further investigations. In this regard, the possible role of many small open reading frames (smORFs) in actinomycete morpho-physiological differentiation is still elusive. In , inactivation of the smORF (SCO2038) - whose product modulates L-tryptophan biosynthesis - impairs production of antibiotics and morphological differentiation. Indeed, it was demonstrated that TrpM is able to interact with PepA (SCO2179), a putative cytosol aminopeptidase playing a key role in antibiotic production and sporulation. In this work, a knock-in (Sco-KI) mutant strain was generated by cloning into overexpressing vector to further investigate the role of in actinomycete growth and morpho-physiological differentiation. Results highlighted that : (i) stimulates growth and actinorhodin (ACT) production; (ii) decreases calcium-dependent antibiotic (CDA) production; (iii) has no effect on undecylprodigiosin production. Metabolic pathways influenced by knock-in were investigated by combining two-difference in gel electrophoresis/nanoliquid chromatography coupled to electrospray linear ion trap tandem mass spectrometry (2D-DIGE/nanoLC-ESI-LIT-MS/MS) and by LC-ESI-MS/MS procedures, respectively. These analyses demonstrated that over-expression of causes an over-representation of factors involved in protein synthesis and nucleotide metabolism as well as a down-representation of proteins involved in central carbon and amino acid metabolism. At the metabolic level, this corresponded to a differential accumulation pattern of different amino acids - including aromatic ones but tryptophan - and central carbon intermediates. PepA was also down-represented in Sco-KI. The latter was produced as recombinant His-tagged protein and was originally proven having the predicted aminopeptidase activity. Altogether, these results highlight the stimulatory effect of in growth and ACT biosynthesis, which are elicited through the modulation of various metabolic pathways and PepA representation, further confirming the complexity of regulatory networks that control antibiotic production in actinomycetes.
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http://dx.doi.org/10.3389/fmicb.2020.00224DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7042404PMC
February 2020

Tuberomics: a molecular profiling for the adaption of edible fungi (Tuber magnatum Pico) to different natural environments.

BMC Genomics 2020 Jan 29;21(1):90. Epub 2020 Jan 29.

A.R.E.A. Foundation, via Tavoleria 28, 56125, Pisa, Italy.

Background: Truffles are symbiotic fungi that develop underground in association with plant roots, forming ectomycorrhizae. They are primarily known for the organoleptic qualities of their hypogeous fruiting bodies. Primarily, Tuber magnatum Pico is a greatly appreciated truffle species mainly distributed in Italy and Balkans. Its price and features are mostly depending on its geographical origin. However, the genetic variation within T. magnatum has been only partially investigated as well as its adaptation to several environments.

Results: Here, we applied an integrated omic strategy to T. magnatum fruiting bodies collected during several seasons from three different areas located in the North, Center and South of Italy, with the aim to distinguish them according to molecular and biochemical traits and to verify the impact of several environments on these properties. With the proteomic approach based on two-dimensional electrophoresis (2-DE) followed by mass spectrometry, we were able to identify proteins specifically linked to the sample origin. We further associated the proteomic results to an RNA-seq profiling, which confirmed the possibility to differentiate samples according to their source and provided a basis for the detailed analysis of genes involved in sulfur metabolism. Finally, geographical specificities were associated with the set of volatile compounds produced by the fruiting bodies, as quantitatively and qualitatively determined through proton transfer reaction-mass spectrometry (PTR-MS) and gas-chromatography-mass spectrometry (GC-MS). In particular, a partial least squares-discriminant analysis (PLS-DA) model built from the latter data was able to return high confidence predictions of sample source.

Conclusions: Results provide a characterization of white fruiting bodies by a wide range of different molecules, suggesting the role for specific compounds in the responses and adaptation to distinct environments.
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http://dx.doi.org/10.1186/s12864-020-6522-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6988325PMC
January 2020

A multi-approach peptidomic analysis of hen egg white reveals novel putative bioactive molecules.

J Proteomics 2020 03 9;215:103646. Epub 2020 Jan 9.

Proteomics & Mass Spectrometry Laboratory, ISPAAM, National Research Council, 80147 Naples, Italy. Electronic address:

Chicken egg white is a raw material broadly used as additive for the preparation of food and cosmetoceutical products. To describe its molecular properties, various proteomic investigations were performed in the last decade characterizing highly abundant components. No peptidomic counterparts were accomplished so far; scientific literature only reports on the characterization of specific bioactive peptides or preparations from egg white and its hydrolysates, which was performed through dedicated functional assays. In this study, a broad description of the egg white peptidome at 24, 336 and 672 h after laying was achieved using three peptide extraction procedures, which were combined with MALDI-TOF-TOF-MS and nanoLC-ESI-Q-Orbitrap-MS/MS analyses. In the whole, 506 peptides were characterized; they mostly resulted from the physiological degradation of intact proteins following the activity of endoprotease ArgC-, trypsin- and plasmin-like enzymes. Eventual detection of peptide post-translational modifications also provided structural information on parental proteins. When analyzed by bioinformatics and/or compared with literature data, identified peptides allowed recognizing a number of protein fragments associated with different hypothetical biological activities. These results confirmed previous observations regarding functional characteristics of egg white unfractionated preparations or purified molecules, emphasizing the useful application of this raw material in human nutrition and cosmetics. Finally, a comparative label-free peptidomic evaluation of samples stored for different times under refrigeration identified 31 peptides showing significant quantitative changes during storage. BIOLOGICAL SIGNIFICANCE: This study provided the largest inventory of peptides described in chicken egg while so far. In addition, it identified a number of protein fragments associated with hypothetical antihypertensive, antioxidant, antiinflammatory, antimicrobial, anticancer, antiviral, antibiofilm, calcium-binding, antidiabetic, antithrombotic, adipogenic differentiating, stimulating/immunostimulating, hormonal, lipid-binding and cell adhesion-affecting activities. These results confirmed previous observations regarding functional characteristics of egg white unfractionated preparations or purified molecules, emphasizing the useful application of this raw material in human nutrition and cosmetics.
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http://dx.doi.org/10.1016/j.jprot.2020.103646DOI Listing
March 2020

Architecture of The Human Ape1 Interactome Defines Novel Cancers Signatures.

Sci Rep 2020 01 8;10(1):28. Epub 2020 Jan 8.

Bioinformatics Core Facility, Centre for Integrative Biology (CIBIO), University of Trento, via Sommarive 18, 38123, Povo (Trento), Italy.

APE1 is essential in cancer cells due to its central role in the Base Excision Repair pathway of DNA lesions and in the transcriptional regulation of genes involved in tumor progression/chemoresistance. Indeed, APE1 overexpression correlates with chemoresistance in more aggressive cancers, and APE1 protein-protein interactions (PPIs) specifically modulate different protein functions in cancer cells. Although important, a detailed investigation on the nature and function of protein interactors regulating APE1 role in tumor progression and chemoresistance is still lacking. The present work was aimed at analyzing the APE1-PPI network with the goal of defining bad prognosis signatures through systematic bioinformatics analysis. By using a well-characterized HeLa cell model stably expressing a flagged APE1 form, which was subjected to extensive proteomics analyses for immunocaptured complexes from different subcellular compartments, we here demonstrate that APE1 is a central hub connecting different subnetworks largely composed of proteins belonging to cancer-associated communities and/or involved in RNA- and DNA-metabolism. When we performed survival analysis in real cancer datasets, we observed that more than 80% of these APE1-PPI network elements is associated with bad prognosis. Our findings, which are hypothesis generating, strongly support the possibility to infer APE1-interactomic signatures associated with bad prognosis of different cancers; they will be of general interest for the future definition of novel predictive disease biomarkers. Future studies will be needed to assess the function of APE1 in the protein complexes we discovered. Data are available via ProteomeXchange with identifier PXD013368.
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http://dx.doi.org/10.1038/s41598-019-56981-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6949240PMC
January 2020

Proteome Alterations in Equine Osteochondrotic Chondrocytes.

Int J Mol Sci 2019 Dec 7;20(24). Epub 2019 Dec 7.

ISPAAM, National Research Council, 80147 Naples, Italy.

Osteochondrosis is a failure of the endochondral ossification that affects developing joints in humans and several animal species. It is a localized idiopathic joint disorder characterized by focal chondronecrosis and growing cartilage retention, which can lead to the formation of fissures, subchondral bone cysts, or intra-articular fragments. Osteochondrosis is a complex multifactorial disease associated with extracellular matrix alterations and failure in chondrocyte differentiation, mainly due to genetic, biochemical, and nutritional factors, as well as traumas. This study describes the main proteomic alterations occurring in chondrocytes isolated from osteochondrotic cartilage fragments. A comparative analysis performed on equine osteochondrotic and healthy chondrocytes showed 26 protein species as differentially represented. In particular, quantitative changes in the extracellular matrix, cytoskeletal and chaperone proteins, and in cell adhesion and signaling molecules were observed in osteochondrotic cells, compared to healthy controls. Functional group analysis annotated most of these proteins in "growth plate and cartilage development", while others were included in "glycolysis and gluconeogenesis", "positive regulation of protein import", "cell-cell adhesion mediator activity", and "mitochondrion nucleoid". These results may help to clarify some chondrocyte functional alterations that may play a significant role in determining the onset and progression of equine osteochondrosis and, being related, of human juvenile osteochondrosis.
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http://dx.doi.org/10.3390/ijms20246179DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6940994PMC
December 2019

Amending an As/Pb contaminated soil with biochar, compost and iron grit: effect on Salix viminalis growth, root proteome profiles and metal(loid) accumulation indexes.

Chemosphere 2020 Apr 25;244:125397. Epub 2019 Nov 25.

Department of Biosciences and Territory, University of Molise, 86090, Pesche, IS, Italy. Electronic address:

There is currently a large amount of research being done into the phytoremediation of polluted soils. Plant installation in contaminated soils may require the application of soil amendments, such as biochar, compost and/or iron grit, which can improve the soil conditions and reduce the metal (loid) phytoavailability and mobility. The beneficial effects of these amendments on soil properties, plant growth and metal (loid) accumulation ability have already been described, although their effect on the plants response machinery has been poorly studied. This study aimed to assess the effect of these amendments on Salix viminalis growth and metal (loid) accumulation, as well as elucidating associated molecular mechanisms. The results showed that the amendment applications improved plant growth by three fold, except for the biochar plus iron combination. It also revealed that metal (loid)s were not effectively translocated from the roots to the shoots (translocation factors <1), their bioaccumulation peaked in the roots, and increased in the presence of iron-based amendments. Corresponding proteomic profiles revealed 34 protein spots differentially represented and suggested that plants counteracted metal (loid)-induced oxidative stress after the addition of biochar and/or compost by eliciting proper defense and signaling pathways, and by redirecting the metabolic fluxes towards primary and secondary metabolism. However, they did highlight the occurrence of oxidative stress markers when the biochar plus iron amendment was applied, which could be both the cause and result of protein degradation impairment.
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http://dx.doi.org/10.1016/j.chemosphere.2019.125397DOI Listing
April 2020

Dehydrogenase/reductase activity of human carbonyl reductase 1 with NADP(H) acting as a prosthetic group.

Biochem Biophys Res Commun 2020 01 20;522(1):259-263. Epub 2019 Nov 20.

University of Pisa, Department of Biology, Biochemistry Unit, Via S. Zeno, 51, Pisa, Italy; Interdepartmental Research Center Nutrafood ''Nutraceuticals and Food for Health'', University of Pisa, Pisa, Italy. Electronic address:

Carbonyl reductase 1 (CBR1) is an NADP-dependent enzyme that exerts a detoxifying role, which catalyses the transformation of carbonyl-containing compounds. The ability of CBR1 to act on adducts between glutathione and lipid peroxidation derived aldehydes has recently been reported. In the present study, exploiting mass spectrometry and fluorescence spectroscopy, evidence is shown that CBR1 is able to retain NADP(H) at the active site even after extensive dialysis, and that this retention may also occur when the enzyme is performing catalysis. This property, together with the multi-substrate specificity of CBR1 in both directions of red/ox reactions, generates inter-conversion red/ox cycles. This particular feature of CBR1, in the case of the transformation of 3-glutathionyl, 4-hydroxynonanal (GSHNE), which is a key substrate of the enzyme in detoxification, supports the disproportionation reaction of GSHNE without any apparent exchange of the cofactor with the solution. The importance of the cofactor as a prosthetic group for other dehydrogenases exerting a detoxification role is discussed.
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http://dx.doi.org/10.1016/j.bbrc.2019.11.090DOI Listing
January 2020