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Identification of Small Molecule Inhibitors against Staphylococcus aureus Dihydroorotase via HTS

journal contribution
posted on 06.05.2022, 17:22 by Amy J Rice, Russell PesaventoRussell Pesavento, Jinhong Ren, Isoo Youn, Youngjin Kwon, Kassapa Ellepola, Chun-Tao CheChun-Tao Che, Michael JohnsonMichael Johnson, Hyun LeeHyun Lee
Drug-resistant Staphylococcus aureus is an imminent threat to public health, increasing the importance of drug discovery utilizing unexplored bacterial pathways and enzyme targets. De novo pyrimidine biosynthesis is a specialized, highly conserved pathway implicated in both the survival and virulence of several clinically relevant pathogens. Class I dihydroorotase (DHOase) is a separate and distinct enzyme present in gram positive bacteria (i.e., S. aureus, B. anthracis) that converts carbamoyl-aspartate (Ca-asp) to dihydroorotate (DHO)-an integral step in the de novo pyrimidine biosynthesis pathway. This study sets forth a high-throughput screening (HTS) of 3000 fragment compounds by a colorimetry-based enzymatic assay as a primary screen, identifying small molecule inhibitors of S. aureus DHOase (SaDHOase), followed by hit validation with a direct binding analysis using surface plasmon resonance (SPR). Competition SPR studies of six hit compounds and eight additional analogs with the substrate Ca-asp determined the best compound to be a competitive inhibitor with a KD value of 11 µM, which is 10-fold tighter than Ca-asp. Preliminary structure-activity relationship (SAR) provides the foundation for further structure-based antimicrobial inhibitor design against S. aureus.

Funding

Multidisciplinary Oral Sciences Training Program | Funder: National Institutes of Health (National Institute of Dental and Craniofacial Research) | Grant ID: T32DE018381

History

Citation

Rice, A. J., Pesavento, R. P., Ren, J., Youn, I., Kwon, Y., Ellepola, K., Che, C. -T., Johnson, M. E.Lee, H. (2021). Identification of Small Molecule Inhibitors against Staphylococcus aureus Dihydroorotase via HTS. International Journal of Molecular Sciences, 22(18), 9984-. https://doi.org/10.3390/ijms22189984

Publisher

MDPI AG

Language

en

issn

1661-6596