TitleCSGID Solves Structures and Identifies Phenotypes for Five Enzymes in .
Publication TypeJournal Article
Year of Publication2018
AuthorsLykins, JD, Filippova, EV, Halavaty, AS, Minasov, G, Zhou, Y, Dubrovska, I, Flores, KJ, Shuvalova, LA, Ruan, J, Bissati, KEl, Dovgin, S, Roberts, CW, Woods, S, Moulton, JD, Moulton, HM, McPhillie, MJ, Muench, SP, Fishwick, CWG, Sabini, E, Shanmugam, D, Roos, DS, McLeod, R, Anderson, WF, Ngô, HM
JournalFront Cell Infect Microbiol
Date Published2018
KeywordsCrystallography, X-Ray, Enzymes, Gene Knockdown Techniques, Models, Molecular, Protein Conformation, Protozoan Proteins, Toxoplasma

, an Apicomplexan parasite, causes significant morbidity and mortality, including severe disease in immunocompromised hosts and devastating congenital disease, with no effective treatment for the bradyzoite stage. To address this, we used the Tropical Disease Research database, crystallography, molecular modeling, and antisense to identify and characterize a range of potential therapeutic targets for toxoplasmosis. Phosphoglycerate mutase II (PGMII), nucleoside diphosphate kinase (NDK), ribulose phosphate 3-epimerase (RPE), ribose-5-phosphate isomerase (RPI), and ornithine aminotransferase (OAT) were structurally characterized. Crystallography revealed insights into the overall structure, protein oligomeric states and molecular details of active sites important for ligand recognition. Literature and molecular modeling suggested potential inhibitors and druggability. The targets were further studied with vivoPMO to interrupt enzyme synthesis, identifying the targets as potentially important to parasitic replication and, therefore, of therapeutic interest. Targeted vivoPMO resulted in statistically significant perturbation of parasite replication without concomitant host cell toxicity, consistent with a previous CRISPR/Cas9 screen showing PGM, RPE, and RPI contribute to parasite fitness. PGM, RPE, and RPI have the greatest promise for affecting replication in tachyzoites. These targets are shared between other medically important parasites and may have wider therapeutic potential.

Alternate JournalFront Cell Infect Microbiol
PubMed ID30345257
PubMed Central IDPMC6182094
Grant ListHHSN272200700058C / AI / NIAID NIH HHS / United States
HHSN272201200026C / AI / NIAID NIH HHS / United States
HHSN272201700060C / AI / NIAID NIH HHS / United States
T35 DK062719 / DK / NIDDK NIH HHS / United States