Citas bibligráficas
Bustillos, E., Vega, E. (2023). Evaluación de la actividad inhibitoria de péptidos sintéticos seleccionados sobre PknG de Mycobacterium tuberculosis [Tesis, Universidad Peruana de Ciencias Aplicadas (UPC)]. http://hdl.handle.net/10757/667385
Bustillos, E., Vega, E. Evaluación de la actividad inhibitoria de péptidos sintéticos seleccionados sobre PknG de Mycobacterium tuberculosis [Tesis]. PE: Universidad Peruana de Ciencias Aplicadas (UPC); 2023. http://hdl.handle.net/10757/667385
@misc{renati/1293866,
title = "Evaluación de la actividad inhibitoria de péptidos sintéticos seleccionados sobre PknG de Mycobacterium tuberculosis",
author = "Vega Loayza, Estefany del Pilar Isabel",
publisher = "Universidad Peruana de Ciencias Aplicadas (UPC)",
year = "2023"
}
Introduction: The identification of new therapeutic targets and antimicrobial compounds are key against drug-resistant tuberculosis. PknG is a virulence factor of Mycobacterium tuberculosis associated with intracellular survival and latency, which is considered a promising target. Objectives: To evaluate the inhibitory activity of selected synthetic peptides for PknG through a luminescence assay. Methods: Recombinant PknG and GarA proteins were expressed and purified by affinity chromatography. PknG phosphorylation reaction and inhibition were assessed by a luminescence assay. For the standardization of the phosphorylation reaction, the composition of the buffer, type of divalent ion as cofactor, and reaction time were evaluated. As inhibition control, AX20017 was used. The percentage inhibition for the candidate peptides was determined. Results: Aliquots of PknG and GarA were obtained at a concentration of 145 µM and 231 µM, respectively. The phosphorylation reaction was standardized to 181.2 nM PknG, 7.7 µM GarA and 10 µM ATP, using PR1 buffer (25 mM HEPES pH 7.5, 100 mM NaCl, 1 mM DTT, 5 mM MgCl2). AX20017 reported an IC50 of 6.71 µM. Six of the selected peptides demonstrated an inhibition percentage higher than 30%. Of these, UPC_07, UPC_08 and UPC_09 showed values close to 39%. Conclusions: An in vitro luminescence assay was standardized to evaluate the inhibition of PknG kinase activity on GarA. Three peptides showed inhibition percentages higher than 38%.
Este ítem está sujeto a una licencia Creative Commons Licencia Creative Commons