Caracterização biofísico-química da proteína de matriz do vírus sincicial respiratório humano.
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Data
2021-06-16
Autores
Guimaraes, Giovana Cavenaghi
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O Vírus Sincicial Respiratório Humano (hRSV) é o principal agente causador de infecções no trato respiratório inferior em crianças e idosos por todo o mundo. A proteína de matriz (M) é essencial para o ciclo viral, uma vez que atua no processo de replicaçãoo, interagindo com proteínas do complexo ribonucleoproteico (RNPs). Posteriormente, a proteína M atua coordenando a transferência dos RNPs para locais de jangadas lipídicas, a partir da interação com o citoesqueleto celular, auxiliando assim no brotamento e liberação de vírions. Sabe-se então, que a proteína atua na célula hospedeira e tem grande importância na replicação viral, desta forma, o objetivo do presente trabalho foi compreender as mudanças conformacionais que podem ocorrer na proteína em diferentes condições de pH, temperatura e salinidade. Utilizando a proteína de matriz recombinante, foram realizados experimentos de fluorescência, dicroísmo circular e calorimetria de varredura diferencial. Os resultados do presente trabalho demonstraram que o processo de desnaturação térmica da proteína é irreversível e apresenta temperatura de desenovelamento de aproximadamente 44 °C. Além disso, sua estrutura secundária é influenciada pelo pH. Porém, mesmo em condição extremamente ácida, a proteína ainda possui parte de sua estrutura preservada. Por fim, o aumento da concentração iônica leva a uma desestabilização da proteína, resultando em diminuição da sua temperatura de desenovelamento. Desta forma os resultados do presente estudo auxiliaram para uma melhor compreensão de características conformacionais da proteína de matriz do hRSV.
The human Respiratory Syncytial Virus (hRSV) is the main cause of infections in the lower respiratory tract in children and elderly all over the world. The matrix protein (M) is essential for the viral cycle once it acts on the replication process interacting with the ribonucleoprotein complex (RNPs). Posteriorly, the M protein interacts with the cellular cytoskeleton coordinating the RNPs transference to lipid rafts aiding the viral budding process and virion release. It is known that the protein acts in the host cell and has a great importance in the viral replication, thus, the objective of this work was to understand the conformational changes that can occur in the protein structure at different conditions of pH, temperature and salinity. Using the recombinant matrix protein, fluorescence, CD and DSC experiments were performed. The results of the present work demonstrates that the denaturation process is irreversible, with a melting temperature of about 44 °C. In addition, the secondary structure of the protein is influenced by the pH, but, even in extremely acid conditions, there is still some conserved secondary structure of the protein. Lastly, the increase of the ionic concentration leads to protein destabilization, resulting in a decrease of the melting temperature. Thus the results of the present work helped to better understand the conformational characteristics of the hRSV matrix protein.
The human Respiratory Syncytial Virus (hRSV) is the main cause of infections in the lower respiratory tract in children and elderly all over the world. The matrix protein (M) is essential for the viral cycle once it acts on the replication process interacting with the ribonucleoprotein complex (RNPs). Posteriorly, the M protein interacts with the cellular cytoskeleton coordinating the RNPs transference to lipid rafts aiding the viral budding process and virion release. It is known that the protein acts in the host cell and has a great importance in the viral replication, thus, the objective of this work was to understand the conformational changes that can occur in the protein structure at different conditions of pH, temperature and salinity. Using the recombinant matrix protein, fluorescence, CD and DSC experiments were performed. The results of the present work demonstrates that the denaturation process is irreversible, with a melting temperature of about 44 °C. In addition, the secondary structure of the protein is influenced by the pH, but, even in extremely acid conditions, there is still some conserved secondary structure of the protein. Lastly, the increase of the ionic concentration leads to protein destabilization, resulting in a decrease of the melting temperature. Thus the results of the present work helped to better understand the conformational characteristics of the hRSV matrix protein.
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Palavras-chave
Proteína M, hRSV, Temperatura, pH, Salinidade, M protein, Temperature, Salinity