Structural and immunological characterization of a new nucleotidyltransferase-like antigen from Paracoccidioides brasiliensis.

Pb27 antigen is an interesting alternative to immunological diagnosis of Paracoccidioidomycosis (PCM) and has demonstrated to be protective in experimental PCM. Its tertiary structure and possible function remained unknown till now. To study Pb27 at the atomic level, the recombinant protein was expressed in Escherichia coli BL21(DE3), purified, and its three-dimensional structure was solved by X-ray crystallography. Based on this structure, we performed a residue correlation analysis and in silico ligand search assays to address a possible biological function to Pb27. We identified Pb27 as a member of the extensive nucleotidyltransferase superfamily. The protein has an αßαßαß topology with two domains (N- and C-terminal domains) and adopts a monomeric form as its biological unit in solution. Structural comparisons with similar members of the superfamily clearly indicate Pb27 C-terminal domain is singular and may play an important role in its biological function. Bioinformatics analysis suggested that Pb27 might bind to ATP and CTP. This suggestion is corroborated by the fact that a magnesium cation is coordinated by two aspartic acid residues present at the active site (between N- and C-terminal domains), as evidenced by X-ray diffraction data. Besides, NMR assays (1H-15N HSQC spectra) confirmed the binding of CTP to Pb27, demonstrating for the first time an interaction between a nucleotide and this protein. Moreover, we evaluated the reactivity of sera from patients with Paracoccidioides brasiliensis infection against the recombinant form of Pb27 and showed that it was recognized by sera from infected and treated patients. Predicted B and T cell epitopes were synthesized and further evaluated against sera of PCM patients, providing information of the most reactive peptides in Pb27 primary structure which interact with specific Pb27 antibodies.

Estudo de estabilidade da eletrossíntese e do desempenho eletroanalítico do compósito poli(l-metionina)-nano-Au na presença de dopamina e de ácido úrico / The stability study of the electrochemical synthesis and electroanalytical function of poly(l-methionine)-nano-Au composite in the presence of dopamine and uric acid

A eletrossíntese e o desempenho eletroanalítico do compósito poli-L-metionina -nano Au foram descritos matematicamente, sendo a primeira descrita etapa por etapa. Os modelos matemáticos foram analisados por meio da teoria de estabilidade linear e da análise de bifurcações. A partir da análise foram inferidas as condições do estado estacionário estável (do modo mais confortável para a eletropolimerização, e do melhor desempenho do sensor), bem como das instabilidades oscilatória e monotônica. As conclusões da modelagem vão ao encontro do observado experimentalmente, completando também o conhecimento do processo.

The electrosynthesis and the electroanalytic function of the poly-L-methioninenano-Au composite were mathematically described and the first one of the processes was described stage by stage. The mathematical models were analyzed by linear stability theory and bifurcation analysis. Basing on the analysis, the stable steady-state conditions, like also oscillatory and monotonic instability conditions were inferred.

Modelagem matemática do desempenho de elétrodos, modificados por camada de ferroceno, nos processos da eletrooxidação de compostos orgânicos em meios neutro e ácido / Modelación matemática del comportamiento de electrodos, modificado por los procesos de la capa de ferroceno en la electro-oxidación de los compuestos orgánicos en medios neutros y ácidos

O comportamento de elétrodos modificados por camada de ferroceno nos processos da eletrooxidação de certos compostos orgânicos em meios neutro e ácido foi descrito matematicamente, sendo os modelos correspondentes analisados por meio da teoria de estabilidade linear e da análise de bifurcações. Através da comparação dos modelos foi avaliada a influência do pH ao comportamento dos elétrodos, modificados por ferroceno.

The behavior of the electrodes modified by ferrocene layer in the electrooxidation of certain organic compounds in neutral and acid media was mathematically described and the mathematical models were analyzed by linear stability theory and bifurcation analysis. Comparing the models, the influence of pH on the electrodes´ behavior was evaluated.