Transition state analogues rescue ribosomes from saporin-L1 ribosome inactivating protein

Matthew B. Sturm, Peter C. Tyler, Gary B. Evans, Vern L. Schramm

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

Ribosome inactivating proteins (RIPs) catalyze the hydrolytic depurination of one or more adenosine residues from eukaryotic ribosomes. Depurination of the ribosomal sarcin-ricin tetraloop (GAGA) causes inhibition of protein synthesis and cellular death. We characterized the catalytic properties of saporin-L1 from Saponaria officinalis (soapwort) leaves, and it demonstrated robust activity against defined nucleic acid substrates and mammalian ribosomes. Transition state analogue mimics of small oligonucleotide substrates of saporin-L1 are powerful, slow-onset inhibitors when adenosine is replaced with the transition state mimic 9-deazaadenine-9-methylene-N-hydroxypyrrolidine (DADMeA). Linear, cyclic, and stem-loop oligonucleotide inhibitors containing DADMeA and based on the GAGA sarcin-ricin tetraloop gave slow-onset tight-binding inhibition constants (Ki*) of 2.3-8.7 nM under physiological conditions and bind up to 40000-fold tighter than RNA substrates. Saporin-L1 inhibition of rabbit reticulocyte translation was protected by these inhibitors. Transition state analogues of saporin-L1 have potential in cancer therapy that employs saporin-L1-linked immunotoxins.

Original languageEnglish (US)
Pages (from-to)9941-9948
Number of pages8
JournalBiochemistry
Volume48
Issue number41
DOIs
Publication statusPublished - Oct 20 2009

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ASJC Scopus subject areas

  • Biochemistry

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