Small-molecule inhibitor leads of Ribosome-inactivating proteins developed using the doorstop approach

Yuan-Ping Pang, Jewn Giew Park, Shaohua Wang, Anuradha Vummenthala, Rajesh K. Mishra, John E. McLaughlin, Rong Di, Jennifer Nielsen Kahn, Nilgun E. Tumer, Laszlo Janosi, Jon Davis, Charles B. Millard

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

Ribosome-inactivating proteins (RIPs) are toxic because they bind to 28S rRNA and depurinate a specific adenine residue from the α-sarcin/ricin loop (SRL), thereby inhibiting protein synthesis. Shiga-like toxins (Stx1 and Stx2), produced by Escherichia coli, are RIPs that cause outbreaks of foodborne diseases with significant morbidity and mortality. Ricin, produced by the castor bean plant, is another RIP lethal to mammals. Currently, no US Food and Drug Administration-approved vaccines nor therapeutics exist to protect against ricin, Shiga-like toxins, or other RIPs. Development of effective small-molecule RIP inhibitors as therapeutics is challenging because strong electrostatic interactions at the RIP•SRL interface make drug-like molecules ineffective in competing with the rRNA for binding to RIPs. Herein, we report small molecules that show up to 20% cell protection against ricin or Stx2 at a drug concentration of 300 nM. These molecules were discovered using the doorstop approach, a new approach to protein•polynucleotide inhibitors that identifies small molecules as doorstops to prevent an active-site residue of an RIP (e.g., Tyr80 of ricin or Tyr77 of Stx2) from adopting an active conformation thereby blocking the function of the protein rather than contenders in the competition for binding to the RIP. This work offers promising leads for developing RIP therapeutics. The results suggest that the doorstop approach might also be applicable in the development of other protein•polynucleotide inhibitors as antiviral agents such as inhibitors of the Z-DNA binding proteins in poxviruses. This work also calls for careful chemical and biological characterization of drug leads obtained from chemical screens to avoid the identification of irrelevant chemical structures and to avoid the interference caused by direct interactions between the chemicals being screened and the luciferase reporter used in screening assays.

Original languageEnglish (US)
Article numbere17883
JournalPLoS One
Volume6
Issue number3
DOIs
StatePublished - 2011

Fingerprint

Ribosome Inactivating Proteins
ribosomes
ricin
Ricin
Molecules
proteins
Shiga-like toxins
Shiga Toxins
drugs
Z-DNA
Poxviridae
Z-Form DNA
ribosomal RNA
Pharmaceutical Preparations
antiviral agents
Ricinus
biopharmaceuticals
therapeutics
castor beans
Foodborne Diseases

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Pang, Y-P., Park, J. G., Wang, S., Vummenthala, A., Mishra, R. K., McLaughlin, J. E., ... Millard, C. B. (2011). Small-molecule inhibitor leads of Ribosome-inactivating proteins developed using the doorstop approach. PLoS One, 6(3), [e17883]. https://doi.org/10.1371/journal.pone.0017883

Small-molecule inhibitor leads of Ribosome-inactivating proteins developed using the doorstop approach. / Pang, Yuan-Ping; Park, Jewn Giew; Wang, Shaohua; Vummenthala, Anuradha; Mishra, Rajesh K.; McLaughlin, John E.; Di, Rong; Kahn, Jennifer Nielsen; Tumer, Nilgun E.; Janosi, Laszlo; Davis, Jon; Millard, Charles B.

In: PLoS One, Vol. 6, No. 3, e17883, 2011.

Research output: Contribution to journalArticle

Pang, Y-P, Park, JG, Wang, S, Vummenthala, A, Mishra, RK, McLaughlin, JE, Di, R, Kahn, JN, Tumer, NE, Janosi, L, Davis, J & Millard, CB 2011, 'Small-molecule inhibitor leads of Ribosome-inactivating proteins developed using the doorstop approach', PLoS One, vol. 6, no. 3, e17883. https://doi.org/10.1371/journal.pone.0017883
Pang, Yuan-Ping ; Park, Jewn Giew ; Wang, Shaohua ; Vummenthala, Anuradha ; Mishra, Rajesh K. ; McLaughlin, John E. ; Di, Rong ; Kahn, Jennifer Nielsen ; Tumer, Nilgun E. ; Janosi, Laszlo ; Davis, Jon ; Millard, Charles B. / Small-molecule inhibitor leads of Ribosome-inactivating proteins developed using the doorstop approach. In: PLoS One. 2011 ; Vol. 6, No. 3.
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abstract = "Ribosome-inactivating proteins (RIPs) are toxic because they bind to 28S rRNA and depurinate a specific adenine residue from the α-sarcin/ricin loop (SRL), thereby inhibiting protein synthesis. Shiga-like toxins (Stx1 and Stx2), produced by Escherichia coli, are RIPs that cause outbreaks of foodborne diseases with significant morbidity and mortality. Ricin, produced by the castor bean plant, is another RIP lethal to mammals. Currently, no US Food and Drug Administration-approved vaccines nor therapeutics exist to protect against ricin, Shiga-like toxins, or other RIPs. Development of effective small-molecule RIP inhibitors as therapeutics is challenging because strong electrostatic interactions at the RIP•SRL interface make drug-like molecules ineffective in competing with the rRNA for binding to RIPs. Herein, we report small molecules that show up to 20{\%} cell protection against ricin or Stx2 at a drug concentration of 300 nM. These molecules were discovered using the doorstop approach, a new approach to protein•polynucleotide inhibitors that identifies small molecules as doorstops to prevent an active-site residue of an RIP (e.g., Tyr80 of ricin or Tyr77 of Stx2) from adopting an active conformation thereby blocking the function of the protein rather than contenders in the competition for binding to the RIP. This work offers promising leads for developing RIP therapeutics. The results suggest that the doorstop approach might also be applicable in the development of other protein•polynucleotide inhibitors as antiviral agents such as inhibitors of the Z-DNA binding proteins in poxviruses. This work also calls for careful chemical and biological characterization of drug leads obtained from chemical screens to avoid the identification of irrelevant chemical structures and to avoid the interference caused by direct interactions between the chemicals being screened and the luciferase reporter used in screening assays.",
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