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

9 Citations (Scopus)

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 2011

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ribonucleoside-triphosphate reductase
Lactobacillus leichmannii
Ribonucleotide Reductases
Electron Spin Resonance Spectroscopy
Paramagnetic resonance
Spectrum Analysis
electron paramagnetic resonance
prokaryotes
Spectroscopy
Technology
Exchange coupling
Catalysis
spectroscopy
catalysis
Tensors
Catalytic Domain
tensors
tubes
preparation

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

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

Cite this

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title = "Investigating the intermediates in the reaction of ribonucleoside triphosphate reductase from Lactobacillus leichmannii: An application of HF EPR-RFQ technology",
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.",
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",
author = "Julia Manzerova and Vladimir Krymov and Gerfen, {Gary J.}",
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AU - Krymov, Vladimir

AU - Gerfen, Gary J.

PY - 2011/12

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AB - 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.

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KW - Electron paramagnetic resonance (EPR)

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KW - Homolysis

KW - Lactobacillus leichmannii

KW - Rapid freeze-quench (RFQ)

KW - Ribonucleoside triphosphate reductase (RTPR)

KW - Ribonucleotide reductase

KW - Thiyl radical

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