A ribozyme with Michaelase activity: Synthesis of the substrate precursors

Alexander Eisenführ, Paramjit Arora, Gerhard Sengle, Leo R. Takaoka, James S. Nowick, Michael Famulok

Research output: Contribution to journalArticle

Abstract

The ability to generate RNA molecules that can catalyze complex organic transformations not only facilitates the reconstruction and plausibility of possible prebiotic reaction pathways but is also crucial for elucidating the potential of the application of RNA catalysts in organic syntheses. Iterative RNA selection previously identified a ribozyme that catalyzes the Michael addition of a cysteine thiol to an α,β-unsaturated amide. This reaction is chemically similar to the rate limiting step of the thymidylate synthase reaction, which is the corresponding reaction of a cysteine thiol to the double-bond of the uracil nucleobase. Here we provide a detailed description of the synthesis of the ribozyme substrates and the substrate oligonucleotides used for its characterization and the investigation of the background reaction. We also describe the further characterization of the ribozyme with respect to substrate specificity. We show that the thiol group of the cysteine nucleophile is essential for the reaction to proceed. When substituted for a thiomethyl group, no reaction takes place.

Original languageEnglish (US)
Pages (from-to)235-249
Number of pages15
JournalBioorganic and Medicinal Chemistry
Volume11
Issue number2
DOIs
StatePublished - Jan 17 2003

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Catalytic RNA
Sulfhydryl Compounds
Cysteine
RNA
Substrates
Synthetic Chemistry Techniques
Prebiotics
Thymidylate Synthase
Nucleophiles
Uracil
Substrate Specificity
Amides
Oligonucleotides
Catalysts
Molecules

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Organic Chemistry
  • Drug Discovery
  • Pharmaceutical Science

Cite this

A ribozyme with Michaelase activity : Synthesis of the substrate precursors. / Eisenführ, Alexander; Arora, Paramjit; Sengle, Gerhard; Takaoka, Leo R.; Nowick, James S.; Famulok, Michael.

In: Bioorganic and Medicinal Chemistry, Vol. 11, No. 2, 17.01.2003, p. 235-249.

Research output: Contribution to journalArticle

Eisenführ, Alexander ; Arora, Paramjit ; Sengle, Gerhard ; Takaoka, Leo R. ; Nowick, James S. ; Famulok, Michael. / A ribozyme with Michaelase activity : Synthesis of the substrate precursors. In: Bioorganic and Medicinal Chemistry. 2003 ; Vol. 11, No. 2. pp. 235-249.
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