RESUMO
Tissue exposure to high levels of tyrosine, which is characteristic of an inborn error of metabolism named Tyrosinemia, is related to severe symptoms, including neurological alterations. The clinical manifestations and pathogenesis of tyrosine neurotoxicity can be recapitulated in experimental models in vivo and in vitro. A widely used experimental model to study brain tyrosine damage is the chronic and acute administration of this amino acid in infant rats. Other research groups and we have extensively studied the pathogenic events in the brain structures of rats exposed to high tyrosine levels. Rats administered acutely and chronically with tyrosine presented decreased and inhibition of the essential metabolism enzymes, e.g., Krebs cycle enzymes and mitochondrial respiratory complexes in the brain structures. These alterations induced by tyrosine toxicity were associated with brain oxidative stress, astrocytes, and, ultimately, cognitive impairments. Notably, in vivo data were corroborated by in vitro studies using cerebral regions homogenates incubated with tyrosine excess. Considering metabolism's importance to brain functioning, we hypothesized that mitochondrial and metabolic dysfunctions are closely related to neurological alterations induced by tyrosine neurotoxicity. Herein, we reviewed the main mechanisms associated with tyrosine neurotoxicity in experimental models, emphasizing the role of mitochondrial dysfunction.
Assuntos
Mitocôndrias/efeitos dos fármacos , Síndromes Neurotóxicas/etiologia , Tirosina/toxicidade , Animais , Encéfalo/efeitos dos fármacos , Encéfalo/metabolismo , Encéfalo/patologia , Modelos Animais de Doenças , Metabolismo Energético/efeitos dos fármacos , Humanos , Mitocôndrias/fisiologia , RatosRESUMO
Halogenated amino acids and peptides are an emerging class of disinfection byproducts (DBPs), having been detected in drinking water and in washed food products. However, the toxicological significance of these emerging DBPs remains unclear. In this study, the cytotoxicity of eight halogenated tyrosyl compounds was investigated in Chinese hamster ovary (CHO) cells using real-time cell analysis (RTCA). Dihalogenated tyrosyl compounds are more cytotoxic than their monohalogenated analogues. The cytotoxicity of the dihalogenated compounds is associated with their ability to induce intracellular reactive oxygen species (ROS), suggesting that oxidative stress is an important toxicity pathway of these compounds. Pearson correlation analysis of the cytotoxicity (IC50 values) of these compounds with eight physicochemical parameters showed strong associations with their lipophilicity (logP) and reactivity (polarizability, ELUMO). Finally, cytotoxicity testing of the concentrated extracts of a chloraminated mixture of eight dipeptides with bromide or iodide showed the cytotoxicity of these mixtures in the order: iodinated peptides > brominated peptides ≥ chlorinated peptides. These results demonstrate that halogenated peptide DBPs are toxicologically relevant, and further research is needed to understand the implications of long-term exposure for human health.
Assuntos
Desinfecção , Halogenação , Tirosina/química , Tirosina/toxicidade , Animais , Células CHO , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Cricetulus , Relação Dose-Resposta a Droga , Estrutura Molecular , Espécies Reativas de Oxigênio/análise , Espécies Reativas de Oxigênio/metabolismo , Tirosina/análogos & derivadosRESUMO
Tyrosinemia type II is an autosomal recessive inborn error of metabolism caused by hepatic cytosolic tyrosine aminotransferase deficiency. Importantly, this disease is associated with neurological and developmental abnormalities in many patients. Considering that the mechanisms underlying neurological dysfunction in hypertyrosinemic patients are poorly understood, in the present work we investigated the levels of cytokines - tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), IL-6 and IL-10 - in cerebellum, hippocampus, striatum of young rats exposed to chronic administration of L-tyrosine. In addition, we also investigated the impact of the supplementation with Omega-3 fatty acids (n-3 PUFA) on the rodent model of Tyrosinemia. Notably, previous study demonstrated an association between L-tyrosine toxicity and n-3 PUFA deficiency. Our results showed a significant increase in the levels of pro- and anti-inflammatory cytokines in brain structures when animals were administered with L-tyrosine. Cerebral cortex and striatum seem to be more susceptible to the inflammation induced by tyrosine toxicity. Importantly, n-3 PUFA supplementation attenuated the alterations on cytokines levels induced by tyrosine exposure in brain regions of infant rats. In conclusion, the brain inflammation is also an important process related to tyrosine neurotoxicity observed in the experimental model of Tyrosinemia. Finally, n-3 PUFA supplementation could be considered as a potential neuroprotective adjunctive therapy for Tyrosinemias, especially type II.
Assuntos
Suplementos Nutricionais , Encefalite/induzido quimicamente , Encefalite/tratamento farmacológico , Ácidos Graxos Ômega-3/administração & dosagem , Mediadores da Inflamação/antagonistas & inibidores , Tirosina/toxicidade , Animais , Animais Recém-Nascidos , Esquema de Medicação , Encefalite/metabolismo , Mediadores da Inflamação/metabolismo , Masculino , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Ratos , Ratos Wistar , Tirosina/administração & dosagemRESUMO
Tyrosinemia type II is a genetic disorder characterized by elevated blood levels of the amino acid tyrosine caused by the deficiency of tyrosine aminotransferase enzyme, resulting in neurologic and developmental difficulties in the patients. Although neurological sequelae are common in Tyrosinemia type II patients, the mechanisms involved are still poorly understood. The oxidative stress appears to be, at least in part, responsible for neurological complication in this inborn error metabolism. We observed that an acute injection of tyrosine in rats caused a massive oxidative stress in different brain structures. The glutathione system and superoxide dismutase enzyme are relevant antioxidant strategies of the cells and tissues, including in the brain. Other important point is the strong relation between oxidative damage and inflammatory events. Herein, we investigated the effects of chronic administration of tyrosine in the hippocampus of young rats, with emphasis in the activity of GSH related enzymes and superoxide dismutase enzyme, and the astrocytosis. We observed that rats exposed to high levels of tyrosine presented an increased content of tyrosine, which was associated with an increment in the activity of glutathione peroxidase and glutathione reductase as well as with a diminished activity of superoxide dismutase. This antioxidant imbalance was accompanied by enhanced glial fibrillary acidic protein immunoreactivity, a marker of astrocytes, in the brain area studied. In conclusion, hippocampus astrogliosis is also a characteristic of brain alteration in Tyrosinemia. In addition, the chronic exposition to high levels of tyrosine is associated with an alteration in the activity of fundamental antioxidant enzymes.
Assuntos
Antioxidantes/metabolismo , Astrócitos/metabolismo , Gliose/metabolismo , Hipocampo/metabolismo , Tirosina/metabolismo , Tirosina/toxicidade , Animais , Animais Recém-Nascidos , Astrócitos/efeitos dos fármacos , Astrócitos/patologia , Esquema de Medicação , Gliose/induzido quimicamente , Gliose/patologia , Hipocampo/efeitos dos fármacos , Hipocampo/patologia , Masculino , Ratos , Ratos Wistar , Tirosina/administração & dosagemRESUMO
PQ Birch represents an allergen-specific immunotherapy for the treatment of birch pollinosis. It consists of native birch pollen extract chemically modified with glutaldehyde adsorbed to L-tyrosine in its microcrystalline form with addition of the adjuvant Monophosphoryl Lipid A (MPL®). A nonclinical safety testing strategy was designed based upon interpretation of current legislation and regulatory intelligence and comprised genotoxicity studies (bacterial reverse mutation and Chinese hamster ovary micronucleus assays), a rat repeat dose toxicology study and a rabbit local tolerance study. No safety findings of concern were found. Thus, no evidence of genotoxicity was found. Relatively minor, immunostimulatory effects were seen following repeated subcutaneous dosing (once every 2 weeks for 13 weeks) as reversible increased white cell count (notably neutrophils), increased globulin level (resulting in decreased albumin/globulin [A/G] ratio) and increased fibrinogen, as well as minor dose site reaction in the form of inflammatory cell infiltrate. These findings are likely due to the immunostimulatory nature of MPL® and/or the presence of L-tyrosine within the adjuvanted vaccine. Similar dose site inflammatory changes to the injected formulation were also noted in the rabbit local tolerance study.
Assuntos
Adjuvantes Imunológicos/toxicidade , Betula/imunologia , Imunoterapia/efeitos adversos , Lipídeo A/análogos & derivados , Pólen/imunologia , Tirosina/toxicidade , Animais , Células CHO , Cricetulus , Feminino , Lipídeo A/toxicidade , Masculino , Testes de Mutagenicidade , Coelhos , Ratos Wistar , Rinite Alérgica Sazonal/terapia , Salmonella typhimurium/efeitos dos fármacos , Salmonella typhimurium/crescimento & desenvolvimento , Pele/efeitos dos fármacosRESUMO
Oxidized tyrosine (O-Tyr) products have been detected in commercial food and have been demonstrated to induce liver injury in our previous study, but the precise mechanisms of the impact induced by dietary O-Tyr are still unclear. Kidney plays an important role in the metabolism of protein. Accumulation of O-Tyr products, especially the dityrosine (Dityr) and advanced oxidation protein products (AOPPs), in vivo was shown to be associated with many kidney diseases. Therefore, this study determined whether chronic exposure to dietary O-Tyr impaired renal function in rats. After O-Tyr treatment for 24 weeks, rats exhibited oxidative stress and protein oxidation in the kidneys, accompanied with inflammatory reaction and renal dysfunction. Elevated extracellular matrix (ECM) contents and the histological examination (HE and Masson stain) results indicated renal fibrosis. The Real-time PCR and Western blotting assay showed that O-Tyr activated phosphorylation of JNK/p38 and up-regulated the expression of transforming growth factor-ß1 (TGF-ß1) and Smad 2/3. These results suggest that dietary O-Tyr could induce oxidative stress, inflammation and renal fibrosis through JNK/p38/TGF-ß1 signaling pathway. Dityr (accounting for 22 % of the total O-Tyr material) may be responsible for the O-Tyr-induced injury. This study also provides a modified procedure for separation and purification of Dityr, the main oxidized product in O-Tyr.
Assuntos
Rim/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Fator de Crescimento Transformador beta1/farmacologia , Tirosina/farmacologia , Animais , Fibrose/induzido quimicamente , Rim/metabolismo , Rim/patologia , Masculino , Camundongos Endogâmicos C57BL , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta1/metabolismo , Tirosina/análogos & derivados , Tirosina/metabolismo , Tirosina/toxicidadeRESUMO
Given the proven utility of natriuretic peptides as serum biomarkers of cardiovascular maladaptation and dysfunction in humans and the high cross-species sequence conservation of atrial natriuretic peptides, natriuretic peptides have the potential to serve as translational biomarkers for the identification of cardiotoxic compounds during multiple phases of drug development. This work evaluated and compared the response of N-terminal proatrial natriuretic peptide (NT-proANP) and N-terminal probrain natriuretic peptide (NT-proBNP) in rats during exercise-induced and drug-induced increases in cardiac mass after chronic swimming or daily oral dosing with a peroxisome proliferator-activated receptor γ agonist. Male Sprague-Dawley rats aged 8 to 10 weeks were assigned to control, active control, swimming, or drug-induced cardiac hypertrophy groups. While the relative heart weights from both the swimming and drug-induced cardiac hypertrophy groups were increased 15% after 28 days of dosing, the serum NT-proANP and NT-proBNP values were only increased in association with cardiac hypertrophy caused by compound administration. Serum natriuretic peptide concentrations did not change in response to adaptive physiologic cardiac hypertrophy induced by a 28-day swimming protocol. These data support the use of natriuretic peptides as fluid biomarkers for the distinction between physiological and drug-induced cardiac hypertrophy.
Assuntos
Hipertrofia Ventricular Esquerda/sangue , Hipertrofia Ventricular Esquerda/induzido quimicamente , Peptídeos Natriuréticos/sangue , Função Ventricular Esquerda/fisiologia , Remodelação Ventricular/fisiologia , Animais , Biomarcadores/sangue , Cardiotoxicidade , Diagnóstico Diferencial , Hipertrofia Ventricular Esquerda/fisiopatologia , Masculino , Oxazóis/administração & dosagem , Oxazóis/toxicidade , PPAR gama/agonistas , Ratos Sprague-Dawley , Natação/fisiologia , Tirosina/administração & dosagem , Tirosina/análogos & derivados , Tirosina/toxicidadeRESUMO
Nitration of both protein-bound and free tyrosine by reactive nitrogen species results in the formation of nitrotyrosine (NT). We previously reported that free NT impairs microtubule polymerization and uncouples endothelial nitric oxide synthase (eNOS) function in pulmonary artery endothelial cells (PAEC). Because microtubules modulate mitochondrial function, we hypothesized that increased NT levels during inflammation and oxidative stress will lead to mitochondrial dysfunction in PAEC. PAEC isolated from fetal lambs were exposed to varying concentrations of free NT. At low concentrations (1-10 µM), NT increased nitration of mitochondrial electron transport chain (ETC) protein subunit complexes I-V and state III oxygen consumption. Higher concentrations of NT (50 µM) caused decreased microtubule acetylation, impaired eNOS interactions with mitochondria, and decreased ETC protein levels. We also observed increases in heat shock protein-90 nitration, mitochondrial superoxide formation, and fragmentation of mitochondria in PAEC. Our data suggest that free NT accumulation may impair microtubule polymerization and exacerbate reactive oxygen species-induced cell damage by causing mitochondrial dysfunction.
Assuntos
Células Endoteliais/efeitos dos fármacos , Mitocôndrias/efeitos dos fármacos , Artéria Pulmonar/efeitos dos fármacos , Tirosina/análogos & derivados , Acetilação , Animais , Células Cultivadas , DNA Mitocondrial/biossíntese , Relação Dose-Resposta a Droga , Complexo de Proteínas da Cadeia de Transporte de Elétrons/metabolismo , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Idade Gestacional , Proteínas de Choque Térmico HSP90/metabolismo , Microtúbulos/efeitos dos fármacos , Microtúbulos/metabolismo , Mitocôndrias/metabolismo , Mitocôndrias/patologia , Óxido Nítrico Sintase Tipo III/metabolismo , Consumo de Oxigênio/efeitos dos fármacos , Ligação Proteica , Artéria Pulmonar/metabolismo , Artéria Pulmonar/patologia , Ovinos , Superóxidos/metabolismo , Tirosina/toxicidadeRESUMO
Tyrosinemia type II is an inborn error of metabolism caused by a deficiency in hepatic cytosolic aminotransferase. Affected patients usually present a variable degree of mental retardation, which may be related to the level of plasma tyrosine. In the present study we evaluated effect of chronic administration of L-tyrosine on the activities of citrate synthase, malate dehydrogenase, succinate dehydrogenase and complexes I, II, II-III and IV in cerebral cortex, hippocampus and striatum of rats in development. Chronic administration consisted of L-tyrosine (500 mg/kg) or saline injections 12 h apart for 24 days in Wistar rats (7 days old); rats were killed 12 h after last injection. Our results demonstrated that L-tyrosine inhibited the activity of citrate synthase in the hippocampus and striatum, malate dehydrogenase activity was increased in striatum and succinate dehydrogenase, complexes I and II-III activities were inhibited in striatum. However, complex IV activity was increased in hippocampus and inhibited in striatum. By these findings, we suggest that repeated administrations of L-tyrosine cause alterations in energy metabolism, which may be similar to the acute administration in brain of infant rats. Taking together the present findings and evidence from the literature, we hypothesize that energy metabolism impairment could be considered an important pathophysiological mechanism underlying the brain damage observed in patients with tyrosinemia type II.
Assuntos
Química Encefálica/efeitos dos fármacos , Metabolismo Energético/efeitos dos fármacos , Tirosina/toxicidade , Tirosinemias , Animais , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/enzimologia , Citrato (si)-Sintase/análise , Citrato (si)-Sintase/antagonistas & inibidores , Ciclo do Ácido Cítrico/efeitos dos fármacos , Corpo Estriado/efeitos dos fármacos , Corpo Estriado/enzimologia , Modelos Animais de Doenças , Complexo de Proteínas da Cadeia de Transporte de Elétrons/análise , Complexo de Proteínas da Cadeia de Transporte de Elétrons/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Hipocampo/enzimologia , Malato Desidrogenase/análise , Malato Desidrogenase/efeitos dos fármacos , Masculino , Proteínas do Tecido Nervoso/análise , Ratos , Ratos WistarRESUMO
Mutations in the tyrosine aminotransferase gene have been identified to cause tyrosinemia type II which is inherited in an autosomal recessive manner. Studies have demonstrated that an excessive production of ROS can lead to reactions with macromolecules, such as DNA, lipids, and proteins. Considering that the L-tyrosine may promote oxidative stress, the main objective of this study was to investigate the in vivo effects of L-tyrosine on DNA damage determined by the alkaline comet assay, in brain and blood of rats. In our acute protocol, Wistar rats (30 days old) were killed 1 h after a single intraperitoneal L-tyrosine injection (500 mg/kg) or saline. For chronic administration, the animals received two subcutaneous injections of L-tyrosine (500 mg/kg, 12-h intervals) or saline administered for 24 days starting at postnatal day (PD) 7 (last injection at PD 31), 12 h after the last injection, the animals were killed by decapitation. We observed that acute administration of L-tyrosine increased DNA damage frequency and damage index in cerebral cortex and blood when compared to control group. Moreover, we observed that chronic administration of L-tyrosine increased DNA damage frequency and damage index in hippocampus, striatum, cerebral cortex and blood when compared to control group. In conclusion, the present work demonstrated that DNA damage can be encountered in brain from animal models of hypertyrosinemia, DNA alterations may represent a further means to explain neurological dysfunction in this inherited metabolic disorder and to reinforce the role of oxidative stress in the pathophysiology of tyrosinemia type II.
Assuntos
Encéfalo/efeitos dos fármacos , Dano ao DNA , Tirosina/toxicidade , Animais , Ensaio Cometa , Dano ao DNA/fisiologia , Estresse Oxidativo/efeitos dos fármacos , Ratos , Ratos Wistar , Tirosina Transaminase/genética , Tirosinemias/induzido quimicamenteRESUMO
A fundamental component for success in drug discovery is the ability to assemble and screen compounds that encompass a broad swath of biologically relevant chemical-diversity space. Achieving this goal in a natural-products-based setting requires access to a wide range of biologically diverse specimens. For this reason, we introduced a crowdsourcing program in which citizen scientists furnish soil samples from which new microbial isolates are procured. Illustrating the strength of this approach, we obtained a unique fungal metabolite, maximiscin, from a crowdsourced Alaskan soil sample. Maximiscin, which exhibits a putative combination of polyketide synthase (PKS), non-ribosomal peptide synthetase (NRPS), and shikimate pathway components, was identified as an inhibitor of UACC-62 melanoma cells (LC50=0.93 µM). The metabolite also exhibited efficacy in a xenograft mouse model. These results underscore the value of building cooperative relationships between research teams and citizen scientists to enrich drug discovery efforts.
Assuntos
Antineoplásicos/metabolismo , Produtos Biológicos/metabolismo , Fungos/metabolismo , Metionina/metabolismo , Tirosina/metabolismo , Animais , Antineoplásicos/uso terapêutico , Antineoplásicos/toxicidade , Produtos Biológicos/uso terapêutico , Produtos Biológicos/toxicidade , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Técnicas de Cocultura , Cristalografia por Raios X , Avaliação Pré-Clínica de Medicamentos , Humanos , Melanoma/tratamento farmacológico , Metionina/química , Metionina/toxicidade , Camundongos , Conformação Molecular , Peptídeo Sintases/metabolismo , Policetídeos/química , Policetídeos/metabolismo , Pseudomonas/metabolismo , Ácido Chiquímico/química , Ácido Chiquímico/metabolismo , Transplante Heterólogo , Tirosina/química , Tirosina/toxicidadeRESUMO
The obesity hormone leptin has been implicated in breast cancer development. Breast cancer cells express the leptin receptor and are able to synthesize leptin in response to obesity-related stimuli. Furthermore, leptin is a positive regulator of vascular endothelial growth factor (VEGF) and high levels of both proteins are associated with worse prognosis in breast cancer patients. Peroxisome proliferator-activated receptor γ (PPARγ) ligands are therapeutic agents used in patient with Type 2 diabetes and obesity which have recently been studied for their potential anti-tumor effect. Here, we studied if these compounds, ciglitazone and GW1929, can affect the expression of leptin and VEGF in breast cancer cells. In MDA-MB-231 and MCF-7 breast cancer cells, treatment with submolar concentrations of ciglitazone and GW1929 elevated the expression of leptin and VEGF mRNA and protein, and increased cell viability and migration. These effects coincided with increased recruitment of PPARγ to the proximal leptin promoter and decreased association of a transcriptional factor Sp1 with this DNA region.
Assuntos
Benzofenonas/farmacologia , Neoplasias da Mama/metabolismo , Leptina/metabolismo , PPAR gama/agonistas , Tiazolidinedionas/farmacologia , Tirosina/análogos & derivados , Fator A de Crescimento do Endotélio Vascular/metabolismo , Benzofenonas/toxicidade , Sítios de Ligação , Neoplasias da Mama/genética , Neoplasias da Mama/patologia , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Leptina/genética , Ligantes , Células MCF-7 , PPAR gama/metabolismo , Regiões Promotoras Genéticas , RNA Mensageiro/metabolismo , Fator de Transcrição Sp1/metabolismo , Tiazolidinedionas/toxicidade , Tirosina/farmacologia , Tirosina/toxicidade , Fator A de Crescimento do Endotélio Vascular/genéticaRESUMO
An aza-cycloisodityrosine analogue of RA-VII, 3, was designed and synthesized. The key aza-cycloisodityrosine unit was prepared by copper(II)-acetate-mediated intramolecular phenylamine/arylboronic acid coupling of dipeptide followed by connection with the tetrapeptide segment to afford a hexapeptide. Subsequent macrocyclization of the hexapeptide with EDC · HCl and HOOBt under dilute conditions gave 3. Analogue 3 showed significant cytotoxic activity against human promyelocytic leukemia HL-60 cells and human colon carcinoma HCT-116 cells, but its activity was weaker than that of parent peptide RA-VII (1).
Assuntos
Antineoplásicos Fitogênicos/química , Peptídeos Cíclicos/química , Peptídeos/química , Tirosina/análogos & derivados , Antineoplásicos Fitogênicos/síntese química , Antineoplásicos Fitogênicos/toxicidade , Compostos Aza/química , Cristalografia por Raios X , Células HCT116 , Células HL-60 , Humanos , Conformação Molecular , Peptídeos/síntese química , Peptídeos/toxicidade , Peptídeos Cíclicos/síntese química , Peptídeos Cíclicos/toxicidade , Rubia/química , Rubia/metabolismo , Tirosina/síntese química , Tirosina/química , Tirosina/toxicidadeRESUMO
The Kaposi's sarcoma-associated herpesvirus (KSHV) G protein-coupled receptor (vGPCR) is a bona fide signaling molecule that is implicated in KSHV-associated malignancies. Whereas vGPCR activates specific cellular signaling pathways in a chemokine-independent fashion, vGPCR binds a broad spectrum of CC and CXC chemokines, and the roles of chemokines in vGPCR tumorigenesis remain poorly understood. We report here that vGPCR is posttranslationally modified by sulfate groups at tyrosine residues within its N-terminal extracellular domain. A chemokine-binding assay demonstrated that the tyrosine sulfate moieties were critical for vGPCR association with GRO-alpha (an agonist) but not with IP-10 (an inverse agonist). A sulfated peptide corresponding to residues 12 through 33 of vGPCR, but not the unsulfated equivalent, partially inhibited vGPCR association with GRO-alpha. Although the vGPCR variant lacking sulfotyrosines activated downstream signaling pathways, the ability of the unsulfated vGPCR variant to induce tumor growth in nude mice was significantly diminished. Furthermore, the unsulfated vGPCR variant was unable to induce the secretion of proliferative cytokines, some of which serve as vGPCR agonists. This implies that autocrine activation by agonist chemokines is critical for vGPCR tumorigenesis. Indeed, GRO-alpha increased vGPCR-mediated AKT phosphorylation and vGPCR tumorigenesis in a sulfotyrosine-dependent manner. Our findings support the conclusion that autocrine activation triggered by chemokine agonists via sulfotyrosines is necessary for vGPCR tumorigenesis, thereby providing a rationale for future therapeutic design targeting the tumorigenic vGPCR.
Assuntos
Herpesvirus Humano 8/patogenicidade , Receptores Acoplados a Proteínas G/fisiologia , Sarcoma de Kaposi/etiologia , Tirosina/análogos & derivados , Animais , Quimiocina CXCL1/antagonistas & inibidores , Quimiocina CXCL1/fisiologia , Quimiocina CXCL10/fisiologia , Citocinas/biossíntese , Humanos , Camundongos , Células NIH 3T3 , Estrutura Terciária de Proteína , Receptores Acoplados a Proteínas G/antagonistas & inibidores , Receptores Acoplados a Proteínas G/química , Transdução de Sinais , Tirosina/toxicidadeRESUMO
Protein tyrosine dimerization and nitration by biologically relevant oxidants usually depend on the intermediate formation of tyrosyl radical ((*)Tyr). In the case of tyrosine oxidation in proteins associated with hydrophobic biocompartments, the participation of unsaturated fatty acids in the process must be considered since they typically constitute preferential targets for the initial oxidative attack. Thus, we postulate that lipid-derived radicals mediate the one-electron oxidation of tyrosine to (*)Tyr, which can afterward react with another (*)Tyr or with nitrogen dioxide ((*)NO(2)) to yield 3,3'-dityrosine or 3-nitrotyrosine within the hydrophobic structure, respectively. To test this hypothesis, we have studied tyrosine oxidation in saturated and unsaturated fatty acid-containing phosphatidylcholine (PC) liposomes with an incorporated hydrophobic tyrosine analogue BTBE (N-t-BOC l-tyrosine tert-butyl ester) and its relationship with lipid peroxidation promoted by three oxidation systems, namely, peroxynitrite, hemin, and 2,2'-azobis (2-amidinopropane) hydrochloride. In all cases, significant tyrosine (BTBE) oxidation was seen in unsaturated PC liposomes, in a way that was largely decreased at low oxygen concentrations. Tyrosine oxidation levels paralleled those of lipid peroxidation (i.e., malondialdehyde and lipid hydroperoxides), lipid-derived radicals and BTBE phenoxyl radicals were simultaneously detected by electron spin resonance spin trapping, supporting an association between the two processes. Indeed, alpha-tocopherol, a known reactant with lipid peroxyl radicals (LOO(*)), inhibited both tyrosine oxidation and lipid peroxidation induced by all three oxidation systems. Moreover, oxidant-stimulated liposomal oxygen consumption was dose dependently inhibited by BTBE but not by its phenylalanine analogue, BPBE (N-t-BOC l-phenylalanine tert-butyl ester), providing direct evidence for the reaction between LOO(*) and the phenol moiety in BTBE, with an estimated second-order rate constant of 4.8 x 10(3) M(-1) s(-1). In summary, the data presented herein demonstrate that LOO(*) mediates tyrosine oxidation processes in hydrophobic biocompartments and provide a new mechanistic insight to understand protein oxidation and nitration in lipoproteins and biomembranes.
Assuntos
Bicamadas Lipídicas/química , Peróxidos/química , Tirosina/química , Espectroscopia de Ressonância de Spin Eletrônica , Hemina/química , Hemina/toxicidade , Peroxidação de Lipídeos , Dióxido de Nitrogênio/química , Dióxido de Nitrogênio/toxicidade , Oxirredução , Peróxidos/toxicidade , Ácido Peroxinitroso/química , Ácido Peroxinitroso/toxicidade , Multimerização Proteica , Tirosina/análogos & derivados , Tirosina/toxicidade , alfa-Tocoferol/farmacologiaRESUMO
The non-protein amino acid meta-Tyrosine (m-Tyr) is produced in cells under conditions of oxidative stress, and m-Tyr has been shown to be toxic to a broad range of biological systems. However, the mechanism by which m-Tyr damages cells is unclear. In E. coli, the quality control (QC) function of phenyalanyl-tRNA synthetase (PheRS) is required for resistantce to m-Tyr. To determine the mechanism of m-Tyr toxicity, we utilitized a strain of E. coli that expresses a QC-defective PheRS. The global responses of E. coli cells to m-Tyr were assessed by RNA-seq, and >500 genes were differentially expressed after the addition of m-Tyr. The most strongly up-regulated genes are involved in unfolded-protein stress response, and cells exposed to m-Tyr contained large, electron-dense protein aggregates, indicating that m-Tyr destabilized a large fraction of the proteome. Additionally, we observed that amino acid biosynthesis and transport regulons, controlled by ArgR, TrpR, and TyrR, and the stringent-response regulon, controlled by DksA/ppGpp, were differentially expressed. m-Tyr resistant mutants were isolated and found to have altered a promoter to increase expression of the enzymes for Phe production or to have altered transporters, which likely result in less uptake or increased efflux of m-Tyr. These findings indicate that when m-Tyr has passed the QC checkpoint by the PheRS, this toxicity of m-Tyr may result from interfering with amino acid metabolism, destabalizing a large number of proteins, and the formation of protein aggregates.
Assuntos
Proteínas de Bactérias/genética , Proteínas de Escherichia coli/genética , Proteínas Repressoras/genética , Tirosina/metabolismo , Aminoacil-tRNA Sintetases/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Guanosina Tetrafosfato/genética , Estresse Oxidativo/genética , Fenilalanina/genética , Agregados Proteicos/genética , Proteoma/genética , Proteoma/metabolismo , Tirosina/genética , Tirosina/toxicidadeRESUMO
The significance of soil-allelochemical interactions was addressed in this paper through studies conducted with m-tyrosine, an amino acid analogue and a potent plant growth inhibitor, in a series of laboratory assays performed in field soil or growth media. The studies were performed as a basis for further evaluation of m-tyrosine activity in field soils containing living plant roots. Here, we examined the role of common soil amendments, including ammonium nitrate fertilizer and activated carbon, in overcoming plant growth inhibition in soils in a laboratory setting by using lettuce as a sensitive indicator of plant toxicity. The phytotoxicity of m-tyrosine was not influenced significantly by soil N amendment; however, when significant amounts of activated carbon were added to the soil medium, growth inhibition in treated lettuce seedlings was strongly reduced. Soil texture did not influence the bioavailability or activity of m-tyrosine, as activity in high organic growth media was similar to that of sand and soil mixtures. Similar to other purported allelochemicals, soil persistence of m-tyrosine was limited, with a predicted half life of less than 1 day in soil in a controlled laboratory setting. Rapid degradation of this molecule likely was due to microbial activity but degradation did not appear to be influenced significantly by soil N amendment. Given the observed activity of m-tyrosine in soil and growth media on seedling growth, potential may exist for development of m-tyrosine as a soil applied herbicide if formulations can be stabilized under soil conditions.
Assuntos
Bioensaio , Laboratórios , Lactuca/efeitos dos fármacos , Solo , Tirosina/toxicidade , Carbono/química , Carbono/farmacologia , Fertilização , Lactuca/crescimento & desenvolvimento , Nitrogênio/química , Nitrogênio/farmacologia , Feromônios/toxicidade , Fatores de Tempo , Tirosina/químicaRESUMO
In this study we reported the synthesis of L-phenyl alanine (Phe) & L-tyrosine (Tyr) Natural Amino acids coated iron oxide magnetic nanoparticles under one-pot and in situ reaction. Functionalized iron oxide magnetic nanoparticles were characterized by X-ray diffraction (XRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), Vibrating Sample Magnetometer (VSM), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM) and Transmission electron microscopy (TEM) techniques. Cellular toxicity of amino acids coated iron oxide magnetic nanoparticles was also investigated on HEK-293 cell lines. Additionally, a hemolysis test of as prepared magnetic nanoparticles were performed. It was found that the synthesized Phe and Tyr coated magnetic nanoparticles (F@Phe NPs and F@Tyr NPs) were spherical in shape with an average size less than 25 nm, also the saturation magnetization (Ms) of the F@Phe NPs and F@Tyr NPs were about 30.02 and 58.23 emu/g, respectively, which was lower than those of bare Fe3O4. The TGA results show that apart from this weight loss, the coated sample shows a weight loss of 5.48, and 6.88% respectively corresponding to loss of Tyr, and Phe which is coated on the Fe3O4 nanoparticles. At a high concentration, less than 2.92 and 3.13% hemolytic activity were observed for F@Phe NPs and F@Tyr NPs, respectively. The F@Phe NPs and F@Tyr NPs show the possibility of using this nanoparticles in the development of in vitro and in vivo pharmaceutical and biomedical fields due to do not possess a toxic effect, good ζ-potential and related small and narrow size distribution.
Assuntos
Composição de Medicamentos/métodos , Nanopartículas de Magnetita/toxicidade , Fenilalanina/toxicidade , Tirosina/toxicidade , Sobrevivência Celular/efeitos dos fármacos , Eritrócitos , Química Verde/métodos , Células HEK293 , Hemólise/efeitos dos fármacos , Humanos , Nanopartículas de Magnetita/administração & dosagem , Nanopartículas de Magnetita/química , Teste de Materiais/métodos , Tamanho da Partícula , Fenilalanina/administração & dosagem , Fenilalanina/química , Nanomedicina Teranóstica/métodos , Testes de Toxicidade , Tirosina/administração & dosagem , Tirosina/químicaRESUMO
As part of our ongoing interest to identify bioactive chemical entities from marine invertebrates, the Red Sea specimen of the Verongid sponge Aplysinella species was studied. Repeated chromatographic fractionation of the methanolic extract of the sponge and HPLC purification of the cytotoxic fractions led to the isolation and the identification of two new compounds, psammaplysin Z and 19-hydroxypsammaplysin Z (1 and 2), together with the previously reported psammaplysins A (3) and E (4). The structural determination of 1-4 was supported by interpretation of their NMR and high-resolution mass spectra. Psammaplysins A and E displayed cytotoxic activity against MBA-MB-231 and HeLa cell lines with IC50 values down to 0.29 µM. On the other hand, psammaplysin Z and 19-hydroxypsammaplysin Z were moderately cytotoxic, indicating the importance of the terminal amine and 2-(methylene)cyclopent-4-ene-1,3-dione moieties in 3 and 4 for potent cytotoxic activity.
Assuntos
Poríferos/metabolismo , Tirosina/química , Animais , Antineoplásicos/química , Antineoplásicos/toxicidade , Produtos Biológicos/química , Produtos Biológicos/toxicidade , Linhagem Celular Tumoral/efeitos dos fármacos , Células HeLa , Humanos , Oceano Índico , Espectroscopia de Ressonância Magnética , Espectrometria de Massas , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Compostos de Espiro/química , Compostos de Espiro/toxicidade , Tirosina/análogos & derivados , Tirosina/toxicidadeRESUMO
In this study, the combined toxicity of UV/chlorinated products on Escherichia coli (E. coli) was investigated when ibuprofen (IBP) and tyrosine (Tyr) were used as two precursors. The median-effect equation and combined index (CI)-isobologram equation were used to evaluate the combined toxicity of UV/chlorinated products. Results revealed that the UV/chlorinated products originated from binary Tyr and IBP showed a synergism in toxicity on Escherichia coli at low concentration level while it turned into a clear antagonism effect above a fa value of 0.2 in the toxicity trial. The combined toxic effects on E. coli were determined by both the potential toxicity mode of specific disinfection byproducts (DBPs) and the complicated interaction caused by Tyr and IBP. The addition of IBP decreased the yield of N-DBPs generated from Tyr, which dominated the effect of combined toxicity. Even though the antagonism predominated in toxicity effect on E. coli, the synergistic toxicity at low dose levels should be getting attention, which was more close to the natural concentration of N-DBPs in waters.