| Congresso Brasileiro de Microbiologia 2023 | Resumo: 665-1 | ||||
Resumo:Leptospirosis is a zoonotic disease that has been observed worldwide, but it is particularly prevalent in low-income tropical countries, with a mortality rate of 5 to 20% in its most severe forms. The species of Leptospira can persist in the environment for weeks or months, made possible by various factors involving second messengers that regulate the bacterium's entire lifestyle. Proteins with PilZ domains are known to be receptors for c-di-GMP and can be found abundantly in the genome of this spirochete. Nevertheless, non-canonical PilZ are not c-di-GMP receptors but have been described as protein-protein adaptors. Given the ability of some Leptospira species to grow in different environmental conditions and in various tissues in their hosts, it suggests that these bacteria possess different signaling pathways that detect external environmental signals and allow for the modification of the bacterial biochemistry, physiology, and behavior. It can be inferred that c-di-GMP has a high potential to orchestrate various important roles in the life cycle of Leptospira. However, little is known about this type of signaling in this human pathogen of public health relevance. Therefore, the objective of this study is to investigate the protein encoded by the locus tag LIC_12546 and determine if there is an interaction between its PilZ domain and the bacterial second messenger c-di-GMP. To achieve this, gene cloning of LIC_12546 was cloned using the pET28a(+) plasmid. Subsequently, protein expression and purification will be carried out using chromatography steps. After obtaining pure protein, interaction assays will be conducted to verify the binding of c-di-GMP to the PilZ domain of the protein encoded by the locus tag LIC_12546 through biochemical experiments such as circular dichroism, Nuclear Magnetic Resonance (NMR), and Isothermal Titration Calorimetry (ITC). Palavras-chave: c-di-GMP, Leptospira interrogans, leptospirosis, PilZ domain Agência de fomento:Capes | |||||