The antiprion compound 6-aminophenanthridine inhibits the protein folding activity of the ribosome by direct competition

Yanhong Pang, Sriram Kurella, Cécile Voisset, Dibyendu Samanta, Debapriya Banerjee, Ariane Schabe, Chanchal Das Gupta, Hervé Galons, Marc Blondel, Suparna Sanyal

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

Domain V of the 23S/25S/28S rRNA of the large ribosomal subunit constitutes the active center for the protein folding activity of the ribosome (PFAR). Using in vitro transcribed domain V rRNAs from Escherichia coli and Saccharomyces cerevisiae as the folding modulators and human carbonic anhydrase as a model protein, we demonstrate that PFAR is conserved from prokaryotes to eukaryotes. It was shown previously that 6-aminophenanthridine (6AP), an antiprion compound, inhibits PFAR. Here, using UV cross-linking followed by primer extension, we show that the protein substrates and 6AP interact with a common set of nucleotides on domain V of 23S rRNA. Mutations at the interaction sites decreased PFAR and resulted in loss or change of the binding pattern for both the protein substrates and 6AP. Moreover, kinetic analysis of human carbonic anhydrase refolding showed that 6AP decreased the yield of the refolded protein but did not affect the rate of refolding. Thus, we conclude that 6AP competitively occludes the protein substrates from binding to rRNA and thereby inhibits PFAR. Finally, we propose a scheme clarifying the mechanism by which 6AP inhibits PFAR.

Original languageEnglish (US)
Pages (from-to)19081-19089
Number of pages9
JournalJournal of Biological Chemistry
Volume288
Issue number26
DOIs
StatePublished - Jun 28 2013

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Pang, Y., Kurella, S., Voisset, C., Samanta, D., Banerjee, D., Schabe, A., Gupta, C. D., Galons, H., Blondel, M., & Sanyal, S. (2013). The antiprion compound 6-aminophenanthridine inhibits the protein folding activity of the ribosome by direct competition. Journal of Biological Chemistry, 288(26), 19081-19089. https://doi.org/10.1074/jbc.M113.466748