{Reference Type}: Journal Article
{Title}: Expression, purification and characterization of CTP synthase PyrG in Staphylococcusaureus.
{Author}: Liu D;Tian Z;Tusong K;Mamat H;Luo Y;
{Journal}: Protein Expr Purif
{Volume}: 221
{Issue}: 0
{Year}: 2024 Sep 3
{Factor}: 2.025
{DOI}: 10.1016/j.pep.2024.106520
{Abstract}: Staphylococcus aureus (S. aureus) presents a significant challenge in both nosocomial and community settings due to its pathogenicity. The emergence of drug-resistant strains exacerbates S. aureus infections, leading to increased mortality rates. PyrG, a member of the cytidine triphosphate (CTP) synthase family, serves as a crucial therapeutic target against S. aureus due to the pivotal role of CTP in cellular metabolism. However, the structural and mechanistic details of S. aureus PyrG remains unknown. Here, we successfully expressed and purified monomeric PyrG. Mutational experiments were conducted based on the results of molecular docking. Based on the results of the molecular docking, we carried out mutation experiments and found that Q386A dramatically decreased the CTP synthase activity compared to the wild-type protein, while Y54A almost completely abolished the activity. Exposure of S. aureus to the kinase inhibitor crizotinib increased expression of gene pyrG. Our results identify the two key sites on PyrG for the CTP synthase activity, and present PyrG gene expression increased during the treatment of crizotinib, which may eventually provide valuable guidance for the development of new drugs against S. aureus infections.