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1.
Spectrochim Acta A Mol Biomol Spectrosc ; 173: 1001-1006, 2017 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-28029507

RESUMO

The interaction between salicylic acid (SA) and the induced disease-resistant protein OsAAA1 in rice was studied using spectroscopy and molecular docking. Ultraviolet (UV) absorption spectroscopy demonstrated an interaction between OsAAA1 protein and SA. Spectroscopy showed that this interaction was a dynamic quenching process. Synchronous fluorescence spectroscopy (SFS) further revealed that this interaction caused changes in the microenvironment of tyrosine and tryptophan and that the interaction site was closer to the tryptophan residue. The structural model of protein OsAAA1 was determined by homology modeling method, and the molecular docking simulation diagram of OsAAA1 with SA was obtained. These models, in combination with a Ramachandran plot analysis, showed amino acid residues ranging from position 240 to position 420 as the possible site interacting with SA. Among them, Gly389, Lys257 and Glu425 might be three key amino acids that can form hydrogen bonds with SA.


Assuntos
Simulação de Acoplamento Molecular , Oryza/química , Proteínas de Plantas/química , Ácido Salicílico/química , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Ácido Salicílico/metabolismo , Espectrometria de Fluorescência
2.
J Environ Sci Health B ; 49(11): 856-63, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25190560

RESUMO

Vinclozolin, a dicarboximide fungicide, is an endocrine disrupting chemical that competes with an androgenic endocrine disruptor compound. Most research has focused on the epigenetic effect of vinclozolin in humans. In terms of ecotoxicology, understanding the effect of vinclozolin on non-target organisms is important. The expression profile of a comprehensive set of genes in the amphipod Hyalella azteca exposed to vinclozolin was examined. The expressed sequence tags in low-dose vinclozolin-treated and -untreated amphipods were isolated and identified by suppression subtractive hybridization. DNA dot blotting was used to confirm the results and establish a subtracted cDNA library for comparing all differentially expressed sequences with and without vinclozolin treatment. In total, 494 differentially expressed genes, including hemocyanin, heatshock protein, cytochrome, cytochrome oxidase and NADH dehydrogenase were detected. Hemocyanin was the most abundant gene. DNA dot blotting revealed 55 genes with significant differential expression. These genes included larval serum protein 1 alpha, E3 ubiquitin-protein ligase, mitochondrial cytochrome c oxidase, mitochondrial protein, proteasome inhibitor, hemocyanin, zinc-finger-containing protein, mitochondrial NADH-ubiquinone oxidoreductase and epididymal sperm-binding protein. Vinclozolin appears to upregulate stress-related genes and hemocyanin, related to immunity. Moreover, vinclozolin downregulated NADH dehydrogenase, related to respiration. Thus, even a non-lethal concentration of vinclozolin still has an effect at the genetic level in H. azteca and presents a potential risk, especially as it would affect non-target organism hormone metabolism.


Assuntos
Anfípodes/efeitos dos fármacos , Anfípodes/genética , Fungicidas Industriais/toxicidade , Regulação da Expressão Gênica/efeitos dos fármacos , Oxazóis/toxicidade , Anfípodes/metabolismo , Animais , Biblioteca Gênica , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Técnicas de Hibridização Subtrativa
3.
Artigo em Inglês | MEDLINE | ID: mdl-22196797

RESUMO

Interaction between the coiled-coil domain of rice blast resistance protein Pi36 and methyl-jasmonate (MeJA) was studied by fluorescence and UV-vis spectroscopic techniques. The quenching mechanism of fluorescence of MeJA by this domain was discussed to be a static quenching procedure. Fluorescence quenching was explored to measure the number of binding sites n and apparent binding constants K. The thermodynamics parameters ΔH, ΔG, ΔS were also calculated. The results indicate the binding reaction was not entropy-driven but enthalpy-driven, and hydrophobic binding played major role in the interaction. The binding sites of MeJA with the coiled-coil structural domain of rice blast resistance protein Pi36 were found to approach the microenvironment of both Tyr and Trp by the synchronous fluorescence spectrometry. The distance r between donor (the coiled-coil domain of rice blast resistance protein Pi36) and acceptor (MeJA) was obtained according to Förster theory of non-radioactive energy transfer.


Assuntos
Acetatos/metabolismo , Ciclopentanos/metabolismo , Resistência à Doença , Magnaporthe/fisiologia , Oryza/metabolismo , Oxilipinas/metabolismo , Doenças das Plantas/microbiologia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Absorção , Sítios de Ligação , Cinética , Estrutura Terciária de Proteína , Espectrometria de Fluorescência , Espectrofotometria Ultravioleta , Relação Estrutura-Atividade , Temperatura , Termodinâmica
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