11 results on '"Ferreira, Juliana C."'
Search Results
2. Residues in the fructose-binding pocket are required for ketohexokinase-A activity.
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Ferreira, Juliana C., Villanueva, Adrian J., Fadl, Samar, Al Adem, Kenana, Cinviz, Zeynep Nur, Nedyalkova, Lyudmila, Cardoso, Thyago H. S., Andrade, Mario Edson, Saksena, Nitin K., Sensoy, Ozge, and Rabeh, Wael M.
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MOLECULAR dynamics , *CANCER genes , *METABOLIC syndrome , *METASTASIS , *TUMOR proteins - Abstract
Excessive fructose consumption is a primary contributor to the global surges in obesity, cancer, and metabolic syndrome. Fructolysis is not robustly regulated and is initiated by ketohexokinase (KHK). In this study, we determined the crystal structure of KHK-A, one of two human isozymes of KHK, in the apo-state at 1.85 Å resolution, and we investigated the roles of residues in the fructose-binding pocket by mutational analysis. Introducing alanine at D15, N42, or N45 inactivated KHK-A, whereas mutating R141 or K174 reduced activity and thermodynamic stability. Kinetic studies revealed that the R141A and K174A mutations reduced fructose affinity by 2- to 4-fold compared to WT KHK-A, without affecting ATP affinity. Molecular dynamics simulations provided mechanistic insights into the potential roles of the mutated residues in ligand coordination and the maintenance of an open state in one monomer and a closed state in the other. Protein–protein interactome analysis indicated distinct expression patterns and downregulation of partner proteins in different tumor tissues, warranting a reevaluation of KHK’s role in cancer development and progression. The connections between different cancer genes and the KHK signaling pathway suggest that KHK is a potential target for preventing cancer metastasis. This study enhances our understanding of KHK-A’s structure and function and offers valuable insights into potential targets for developing treatments for obesity, cancer, and metabolic syndrome. [ABSTRACT FROM AUTHOR]
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- 2024
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3. Baccharin and p-coumaric acid from green propolis mitigate inflammation by modulating the production of cytokines and eicosanoids
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Ferreira, Juliana C., Reis, Mouzarllem B., Coelho, Giovanna D.P., Gastaldello, Gabriel H., Peti, Ana Paula F., Rodrigues, Débora M., Bastos, Jairo K., Campo, Vanessa L., Sorgi, Carlos A., Faccioli, Lúcia H., Gardinassi, Luiz G., Tefé-Silva, Cristiane, and Zoccal, Karina F.
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- 2021
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4. pH profiles of 3-chymotrypsin-like protease (3CLpro) from SARS-CoV-2 elucidate its catalytic mechanism and a histidine residue critical for activity.
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Al Adem, Kenana, Ferreira, Juliana C., Fadl, Samar, and Rabeh, Wael M.
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COVID-19 , *ENZYME kinetics , *SARS-CoV-2 , *HISTIDINE - Abstract
3-Chymotrypsin-like protease (3CLpro) is a promising drug target for coronavirus disease 2019 and related coronavirus diseases because of the essential role of this protease in processing viral polyproteins after infection. Understanding the detailed catalytic mechanism of 3CLpro is essential for designing effective inhibitors of infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Molecular dynamics studies have suggested pH-dependent conformational changes of 3CLpro, but experimental pH profiles of SARS-CoV-2 3CLpro and analyses of the conserved active-site histidine residues have not been reported. In this work, pHdependence studies of the kinetic parameters of SARS-CoV-2 3CLpro revealed a bell-shaped pH profile with 2 pKa values (6.9 ± 0.1 and 9.4 ± 0.1) attributable to ionization of the catalytic dyad His41 and Cys145, respectively. Our investigation of the roles of conserved active-site histidines showed that different amino acid substitutions of His163 produced inactive enzymes, indicating a key role of His163 in maintaining catalytically active SARS-CoV-2 3CLpro. By contrast, the H164A and H172A mutants retained 75% and 26% of the activity of WT, respectively. The alternative amino acid substitutions H172K and H172R did not recover the enzymatic activity, whereas H172Y restored activity to a level similar to that of the WT enzyme. The pH profiles of H164A, H172A, and H172Y were similar to those of the WT enzyme, with comparable pKa values for the catalytic dyad. Taken together, the experimental data support a general base mechanism of SARS-CoV-2 3CLpro and indicate that the neutral states of the catalytic dyad and active-site histidine residues are required for maximum enzyme activity. [ABSTRACT FROM AUTHOR]
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- 2023
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5. Key dimer interface residues impact the catalytic activity of 3CLpro, the main protease of SARS-CoV-2.
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Ferreira, Juliana C., Fadl, Samar, and Rabeh, Wael M.
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SARS-CoV-2 , *CATALYTIC activity , *VIRUS removal (Water purification) - Abstract
3C-like protease (3CLpro) processes and liberates functional viral proteins essential for the maturation and infectivity of severe acute respiratory syndrome coronavirus 2, the virus responsible for COVID-19. It has been suggested that 3CLpro is catalytically active as a dimer, making the dimerization interface a target for antiviral development. Guided by structural analysis, here we introduced single amino acid substitutions at nine residues at three key sites of the dimer interface to assess their impact on dimerization and activity. We show that at site 1, alanine substitution of S1 or E166 increased by twofold or reduced relative activity, respectively. At site 2, alanine substitution of S10 or E14 eliminated activity, whereas K12A exhibited (60% relative activity. At site 3, alanine substitution of R4, E290, or Q299 eliminated activity, whereas S139A exhibited 46% relative activity. We further found that the oligomerization states of the dimer interface mutants varied; the inactive mutants R4A, R4Q, S10A/C, E14A/ D/Q/S, E290A, and Q299A/E were present as dimers, demonstrating that dimerization is not an indication of catalytically active 3CLpro. In addition, present mostly as monomers, K12A displayed residual activity, which could be attributed to the conspicuous amount of dimer present. Finally, differential scanning calorimetry did not reveal a direct relationship between the thermodynamic stability of mutants with oligomerization or catalytic activity. These results provide insights on two allosteric sites, R4/E290 and S10/E14, that may promote the design of antiviral compounds that target the dimer interface rather than the active site of severe acute respiratory syndrome coronavirus 2 3CLpro. [ABSTRACT FROM AUTHOR]
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- 2022
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6. Linker residues regulate the activity and stability of hexokinase 2, a promising anticancer target.
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Ferreira, Juliana C., Khrbtli, Abdul-Rahman, Shetler, Cameron L., Mansoor, Samman, Ali, Liaqat, Sensoy, Ozge, and Rabeh, Wael M.
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GLUCOKINASE , *GLUCOSE metabolism , *C-terminal residues , *TUMOR growth , *ISOENZYMES - Abstract
Hexokinase (HK) catalyzes the first step in glucose metabolism, making it an exciting target for the inhibition of tumor initiation and progression due to their elevated glucose metabolism. The upregulation of hexokinase-2 (HK2) in many cancers and its limited expression in normal tissues make it a particularly attractive target for the selective inhibition of cancer growth and the eradication of tumors with limited side effects. The design of such safe and effective anticancer therapeutics requires the development of HK2-specific inhibitors that will not interfere with other HK isozymes. As HK2 is unique among HKs in having a catalytically active N-terminal domain (NTD), we have focused our attention on this region. We previously found that NTD activity is affected by the size of the linker helix-α13 that connects the N- and C-terminal domains of HK2. Three nonactive site residues (D447, S449, and K451) at the beginning of the linker helix-α13 have been found to regulate the NTD activity of HK2. Mutation of these residues led to increased dynamics, as shown via hydrogen deuterium exchange analysis and molecular dynamic simulations. D447A contributed the most to the enhanced dynamics of the NTD, with reduced calorimetric enthalpy of HK2. Similar residues exist in the C-terminal domain (CTD) but are unnecessary for HK1 and HK2 activity. Thus, we postulate these residues serve as a regulatory site for HK2 and may provide new directions for the design of anticancer therapeutics that reduce the rate of glycolysis in cancer through specific inhibition of HK2. [ABSTRACT FROM AUTHOR]
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- 2021
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7. Comparison of 3 modes of automated weaning from mechanical ventilation: A bench study.
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Morato, José B., Sakuma, Mayara T. A., Ferreira, Juliana C., and Caruso, Pedro
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MECHANICAL ventilators -- Evaluation ,ANALYSIS of variance ,ARTIFICIAL respiration ,COMPARATIVE studies ,PROBABILITY theory ,STATISTICS ,DATA analysis ,DATA analysis software ,DESCRIPTIVE statistics - Abstract
Purpose: Automated weaning modes are available in some mechanical ventilators, but no studies compared them hitherto. We compared the performance of 3 automated modes under standard and challenging situations. Methods: We used a lung simulator to compare 3 automated modes, adaptive support ventilation (ASV), mandatory rate ventilation (MRV), and Smartcare, in 6 situations, weaning success, weaning failure, weaning success with extreme anxiety, weaning success with Cheyne-Stokes, weaning success with irregular breathing, and weaning failure with ineffective efforts. Results: The 3 modes correctly recognized the situations of weaning success and failure, even when anxiety or irregular breathing were present but incorrectly recognized weaning success with Cheyne- Stokes. MRV incorrectly recognized weaning failure with ineffective efforts. Time to pressure support (PS) stabilization was shorter for ASV (1-2 minutes for all situations) and MRV (1-7 minutes) than for Smartcare (8-78 minutes). ASV had higher rates of PS oscillations per 5 minutes (4-15), compared with Smartcare (0-1) and MRV (0-12), except when extreme anxiety was present. Conclusions: Smartcare, ASV, and MRV were equally able to recognize weaning success and failure, despite the presence of anxiety or irregular breathing but performed incorrectly in the presence of Cheyne-Stokes. PS behavior over the time differs among modes, with ASV showing larger and more frequent PS oscillations over the time. Clinical studies are needed to confirm our results. [ABSTRACT FROM AUTHOR]
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- 2012
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8. Substrate specificity, physicochemical and kinetic properties of a trypsin from the giant Amazonian fish pirarucu (Arapaima gigas).
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de Freitas-Júnior, Augusto Cézar V., da Costa, Helane Maria S., Marcuschi, Marina, Icimoto, Marcelo Y., Machado, Marcelo F.M., Machado, Maurício F.M., Ferreira, Juliana C., de Oliveira, Vitor M.S.B.B., Buarque, Diego S., and Bezerra, Ranilson S.
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TRYPSIN ,ENZYME specificity ,MOLECULAR weights ,SERINE proteinases ,ENZYMES - Abstract
Specificity studies can contribute to the understanding of how substrate binding and catalysis change among similar enzymes. This study evaluated through the substrate specificity, physicochemical and kinetic properties of a trypsin from the pyloric caeca of pirarucu (Arapaima gigas). For such purposes, two series of FRET-peptides (Abz-RXFK-Eddnp and Abz-XRFK-Eddnp) were used. The mature trypsin molecular mass was 23.5 kDa and N-terminal sequence was IVGGYECPRNSVPYQVSLNVGYH. Moreover, higher trypsin catalytic efficiency was observed for the substrates containing Arg, Tyr and Ser at P1′ and Lys and Val at P2. However, catalytic deficiency was found in the presence of Pro, Trp, Asp, Gly, Glu, Ala at P1' and Gly, Glu, Trp, Asn, Gln and Tyr at P2. Using z-FR-MCA as a substrate, the optimum pH (9.25) and temperature (45 °C) were determined by pseudo-first-order kinetics. The enzyme retained all of its initial activity after 180 min incubation at temperatures up to 45 °C. Kinetic assays in the presence of Ca
2+ showed an increase of approximately 7-fold in enzyme catalytic efficiency (k cat/ K m). On the other hand, Na+ , K+ and Mg2+ were able to inhibit pirarucu trypsin. Therefore, the knowledge of kinetic and biochemical properties, as well as enzyme specificity, are important for the evaluation of biotechnological use of this enzyme and may contribute to the development of more efficient substrates for fish trypsin. [Display omitted] • N-terminal sequence of Arapaima gigas trypsin is homologous to other fish trypsins. • Pirarucu trypsin was able to hydrolyze substrates with Arg, Tyr and Ser at P1'. • Pirarucu trypsin was stable at 45 °C for 180 min using z-FR-MCA. • Calcium activated trypsin, whereas sodium, potassium and magnesium inhibited it. [ABSTRACT FROM AUTHOR]- Published
- 2021
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9. KBE009: An antimalarial bestatin-like inhibitor of the Plasmodium falciparum M1 aminopeptidase discovered in an Ugi multicomponent reaction-derived peptidomimetic library.
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González-Bacerio, Jorge, Maluf, Sarah El Chamy, Méndez, Yanira, Pascual, Isel, Florent, Isabelle, Melo, Pollyana M.S., Budu, Alexandre, Ferreira, Juliana C., Moreno, Ernesto, Carmona, Adriana K., Rivera, Daniel G., Alonso del Rivero, Maday, and Gazarini, Marcos L.
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MALARIA treatment , *ANTIMALARIALS , *PLASMODIUM falciparum , *AMINOPEPTIDASES , *PEPTIDOMIMETICS , *DRUG development - Abstract
Malaria is a global human parasitic disease mainly caused by the protozoon Plasmodium falciparum . Increased parasite resistance to current drugs determines the relevance of finding new treatments against new targets. A novel target is the M1 alanyl-aminopeptidase from P. falciparum (PfA-M1), which is essential for parasite development in human erythrocytes and is inhibited by the pseudo-peptide bestatin. In this work, we used a combinatorial multicomponent approach to produce a library of peptidomimetics and screened it for the inhibition of recombinant PfA-M1 (rPfA-M1) and the in vitro growth of P. falciparum erythrocytic stages (3D7 and FcB1 strains). Dose-response studies with selected compounds allowed identifying the bestatin-based peptidomimetic KBE009 as a submicromolar rPfA-M1 inhibitor ( K i = 0.4 μM) and an in vitro antimalarial compound as potent as bestatin (IC 50 = 18 μM; without promoting erythrocyte lysis). At therapeutic-relevant concentrations, KBE009 is selective for rPfA-M1 over porcine APN (a model of these enzymes from mammals), and is not cytotoxic against HUVEC cells. Docking simulations indicate that this compound binds PfA-M1 without Zn 2+ coordination, establishing mainly hydrophobic interactions and showing a remarkable shape complementarity with the active site of the enzyme. Moreover, KBE009 inhibits the M1-type aminopeptidase activity (Ala-7-amido-4-methylcoumarin substrate) in isolated live parasites with a potency similar to that of the antimalarial activity (IC 50 = 82 μM), strongly suggesting that the antimalarial effect is directly related to the inhibition of the endogenous PfA-M1. These results support the value of this multicomponent strategy to identify PfA-M1 inhibitors, and make KBE009 a promising hit for drug development against malaria. [ABSTRACT FROM AUTHOR]
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- 2017
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10. On the mechanisms of phenothiazine-induced mitochondrial permeability transition: Thiol oxidation, strict Ca2+ dependence, and cyt c release
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Cruz, Thiago S., Faria, Priscila A., Santana, Débora P., Ferreira, Juliana C., Oliveira, Vitor, Nascimento, Otaciro R., Cerchiaro, Giselle, Curti, Carlos, Nantes, Iseli L., and Rodrigues, Tiago
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THIOLS , *PHENOTHIAZINE , *SCHIZOPHRENIA treatment , *ANTIOXIDANTS , *CELL death , *MITOCHONDRIAL membranes , *PEROXIDASE - Abstract
Abstract: Phenothiazines (PTZ) are drugs widely used in the treatment of schizophrenia. Trifluoperazine, a piperazinic PTZ derivative, has been described as inhibitor of the mitochondrial permeability transition (MPT). We reported previously the antioxidant activity of thioridazine at relatively low concentrations associated to the inhibition of the MPT (Brit. J. Pharmacol., 2002;136:136–142). In this study, it was investigated the induction of MPT by PTZ derivatives at concentrations higher than 10μM focusing on the molecular mechanism involved. PTZ promoted a dose–response mitochondrial swelling accompanied by mitochondrial transmembrane potential dissipation and calcium release, being thioridazine the most potent derivative. PTZ-induced MPT was partially inhibited by CsA or Mg2+ and completely abolished by the abstraction of calcium. The oxidation of reduced thiol group of mitochondrial membrane proteins by PTZ was upstream the PTP opening and it was not sufficient to promote the opening of PTP that only occurred when calcium was present in the mitochondrial matrix. EPR experiments using DMPO as spin trapping excluded the participation of reactive oxygen species on the PTZ-induced MPT. Since PTZ give rise to cation radicals chemically by the action of peroxidases and cyanide inhibited the PTZ-induced swelling, we propose that PTZ bury in the inner mitochondrial membrane and the chemically generated PTZ cation radicals modify specific thiol groups that in the presence of Ca2+ result in MPT associated to cytochrome c release. These findings contribute for the understanding of mechanisms of MPT induction and may have implications for the cell death induced by PTZ. [Copyright &y& Elsevier]
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- 2010
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11. Oligomerization of the cysteinyl-rich oligopeptidase EP24.15 is triggered by S-glutathionylation
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Demasi, Marilene, Piassa Filho, Gilberto M., Castro, Leandro M., Ferreira, Juliana C., Rioli, Vanessa, and Ferro, Emer S.
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MAJOR histocompatibility complex , *TRANSPLANTATION immunology , *PHOSPHORUS compounds , *IRON-nickel-phosphorus alloys - Abstract
Abstract: Thimet oligopeptidase (EC 3.4.24.15; EP24.15) is a thiol-rich metallopeptidase ubiquitously distributed in mammalian tissues and involved in oligopeptide metabolism both within and outside cells. Fifteen Cys residues are present in the rat EP24.15 protein, seven are solvent accessible, and two are found inside the catalytic site cleft; no intraprotein disulfide is described. In the present investigation, we show that mammalian immunoprecipitated EP24.15 is S-glutathionylated. In vitro EP24.15 S-glutathionylation was demonstrated by the incubation of bacterial recombinant EP24.15 with oxidized glutathione concentration as low as 10 μM. The in vitro S-glutathionylation of EP24.15 was responsible for its oxidative oligomerization to dimer and trimer complexes. EP24.15 immunoprecipitated from cells submitted to oxidative challenge showed increased trimeric forms and decreased S-glutathionylation compared to immunoprecipitated protein from control cells. Our present data also show that EP24.15 maximal enzymatic activity is maintained by partial S-glutathionylation, a mechanism that apparently regulates the protein oligomerization. Present results raise the possibility of an unconventional property of protein S-glutathionylation, inducing oligomerization by interprotein thiol–disulfide exchange. [Copyright &y& Elsevier]
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- 2008
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