CSGID solves structures and identifies phenotypes for five enzymes in Toxoplasma gondii

Joseph D. Lykins, Ekaterina V. Filippova, Andrei S. Halavaty, George Minasova, Ying Zhou, Ievgeniia Dubrovska, Kristin J. Flores, Ludmilla A. Shuvalova, Jiapeng Ruan, Kamal El Bissati, Sarah Dovgin, Craig W. Roberts, Stuart Woods, Jon D. Moulton, Hong Moulton, Martin J. McPhillie, Stephen P. Muench, Colin W. G. Fishwick, Elisabetta Sabini, Dhanasekaran Shanmugam & 4 others David S. Roos, Rima McLeod, Wayne F. Anderson, Huân M Ngô

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Toxoplasma gondii, 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.
LanguageEnglish
Article number352
Number of pages21
JournalFrontiers in Cellular and Infection Microbiology : Parasite and Host
Volume8
DOIs
Publication statusPublished - 5 Oct 2018

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Toxoplasma
Racemases and Epimerases
Parasites
Phenotype
Crystallography
Phosphates
Enzymes
Clustered Regularly Interspaced Short Palindromic Repeats
Phosphoglycerate Mutase
Ornithine-Oxo-Acid Transaminase
Nucleoside-Diphosphate Kinase
Chemical Databases
Toxoplasmosis
Immunocompromised Host
Catalytic Domain
Therapeutics
Ligands
Morbidity
Mortality
Research

Keywords

  • Toxoplasma gondii
  • phyophoglycerate mutase
  • nucleoside diphoshate kinase

Cite this

Lykins, Joseph D. ; Filippova, Ekaterina V. ; Halavaty, Andrei S. ; Minasova, George ; Zhou, Ying ; Dubrovska, Ievgeniia ; Flores, Kristin J. ; Shuvalova, Ludmilla A. ; Ruan, Jiapeng ; El Bissati, Kamal ; Dovgin, Sarah ; Roberts, Craig W. ; Woods, Stuart ; Moulton, Jon D. ; Moulton, Hong ; McPhillie, Martin J. ; Muench, Stephen P. ; Fishwick, Colin W. G. ; Sabini, Elisabetta ; Shanmugam, Dhanasekaran ; Roos, David S. ; McLeod, Rima ; Anderson, Wayne F. ; Ngô, Huân M. / CSGID solves structures and identifies phenotypes for five enzymes in Toxoplasma gondii. In: Frontiers in Cellular and Infection Microbiology : Parasite and Host. 2018 ; Vol. 8.
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abstract = "Toxoplasma gondii, 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.",
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author = "Lykins, {Joseph D.} and Filippova, {Ekaterina V.} and Halavaty, {Andrei S.} and George Minasova and Ying Zhou and Ievgeniia Dubrovska and Flores, {Kristin J.} and Shuvalova, {Ludmilla A.} and Jiapeng Ruan and {El Bissati}, Kamal and Sarah Dovgin and Roberts, {Craig W.} and Stuart Woods and Moulton, {Jon D.} and Hong Moulton and McPhillie, {Martin J.} and Muench, {Stephen P.} and Fishwick, {Colin W. G.} and Elisabetta Sabini and Dhanasekaran Shanmugam and Roos, {David S.} and Rima McLeod and Anderson, {Wayne F.} and Ng{\^o}, {Hu{\^a}n M}",
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Lykins, JD, Filippova, EV, Halavaty, AS, Minasova, G, Zhou, Y, Dubrovska, I, Flores, KJ, Shuvalova, LA, Ruan, J, El Bissati, K, Dovgin, S, Roberts, CW, Woods, S, Moulton, JD, Moulton, H, McPhillie, MJ, Muench, SP, Fishwick, CWG, Sabini, E, Shanmugam, D, Roos, DS, McLeod, R, Anderson, WF & Ngô, HM 2018, 'CSGID solves structures and identifies phenotypes for five enzymes in Toxoplasma gondii' Frontiers in Cellular and Infection Microbiology : Parasite and Host, vol. 8, 352. https://doi.org/10.3389/fcimb.2018.00352

CSGID solves structures and identifies phenotypes for five enzymes in Toxoplasma gondii. / Lykins, Joseph D.; Filippova, Ekaterina V.; Halavaty, Andrei S.; Minasova, George; Zhou, Ying; Dubrovska, Ievgeniia; Flores, Kristin J.; Shuvalova, Ludmilla A.; Ruan, Jiapeng; El Bissati, Kamal; Dovgin, Sarah; Roberts, Craig W.; Woods, Stuart; Moulton, Jon D.; Moulton, Hong ; McPhillie, Martin J.; Muench, Stephen P.; Fishwick, Colin W. G.; Sabini, Elisabetta ; Shanmugam, Dhanasekaran; Roos, David S.; McLeod, Rima; Anderson, Wayne F.; Ngô, Huân M.

In: Frontiers in Cellular and Infection Microbiology : Parasite and Host, Vol. 8, 352, 05.10.2018.

Research output: Contribution to journalArticle

TY - JOUR

T1 - CSGID solves structures and identifies phenotypes for five enzymes in Toxoplasma gondii

AU - Lykins, Joseph D.

AU - Filippova, Ekaterina V.

AU - Halavaty, Andrei S.

AU - Minasova, George

AU - Zhou, Ying

AU - Dubrovska, Ievgeniia

AU - Flores, Kristin J.

AU - Shuvalova, Ludmilla A.

AU - Ruan, Jiapeng

AU - El Bissati, Kamal

AU - Dovgin, Sarah

AU - Roberts, Craig W.

AU - Woods, Stuart

AU - Moulton, Jon D.

AU - Moulton, Hong

AU - McPhillie, Martin J.

AU - Muench, Stephen P.

AU - Fishwick, Colin W. G.

AU - Sabini, Elisabetta

AU - Shanmugam, Dhanasekaran

AU - Roos, David S.

AU - McLeod, Rima

AU - Anderson, Wayne F.

AU - Ngô, Huân M

PY - 2018/10/5

Y1 - 2018/10/5

N2 - Toxoplasma gondii, 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.

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KW - Toxoplasma gondii

KW - phyophoglycerate mutase

KW - nucleoside diphoshate kinase

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M3 - Article

VL - 8

JO - Frontiers in Cellular and Infection Microbiology : Parasite and Host

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JF - Frontiers in Cellular and Infection Microbiology : Parasite and Host

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ER -