Investigating the intermediates in the reaction of ribonucleoside triphosphate reductase from Lactobacillus leichmannii: An application of HF EPR-RFQ technology

Julia Manzerova, Vladimir Krymov, Gary J. Gerfen

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

In this investigation high-frequency electron paramagnetic resonance spectroscopy (HFEPR) in conjunction with innovative rapid freeze-quench (RFQ) technology is employed to study the exchange-coupled thiyl radical-cob(II)alamin system in ribonucleotide reductase from a prokaryote Lactobacillus leichmannii. The size of the exchange coupling (J ex) and the values of the thiyl radical g tensor are refined, while confirming the previously determined (Gerfen et al. (1996) [20]) distance between the paramagnets. Conclusions relevant to ribonucleotide reductase catalysis and the architecture of the active site are presented. A key part of this work has been the development of a unique RFQ apparatus for the preparation of millisecond quench time RFQ samples which can be packed into small (0.5 mm ID) sample tubes used for CW and pulsed HFEPR - lack of this ability has heretofore precluded such studies. The technology is compatible with a broad range of spectroscopic techniques and can be readily adopted by other laboratories.

Original languageEnglish (US)
Pages (from-to)32-45
Number of pages14
JournalJournal of Magnetic Resonance
Volume213
Issue number1
DOIs
StatePublished - Dec 1 2011

Keywords

  • Active site
  • Adenosylcobalamin
  • Dipolar interaction
  • Electron paramagnetic resonance (EPR)
  • Enzyme
  • Enzymology
  • Exchange interaction (J )
  • Exchange-coupled pair
  • High-frequency (HF)
  • Homolysis
  • Lactobacillus leichmannii
  • Rapid freeze-quench (RFQ)
  • Ribonucleoside triphosphate reductase (RTPR)
  • Ribonucleotide reductase
  • Thiyl radical

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Nuclear and High Energy Physics
  • Condensed Matter Physics

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