Evaluation of spin labels for in-cell EPR by analysis of nitroxide reduction in cell extract of Xenopus laevis oocytes

Azarkh, Mykhailo; Okle, Oliver; Eyring, Philipp; Dietrich, Daniel R.; Drescher, Malte

Evaluation of spin labels for in-cell EPR by analysis of nitroxide reduction in cell extract of Xenopus laevis oocytes

Dateien

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Datum

2011

Open Access-Veröffentlichung

Open Access Green

Sammlungen

Chemie
Biologie
Zukunftskolleg

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Journal of Magnetic Resonance. 2011, 212(2), pp. 450-454. ISSN 1090-7807. eISSN 1096-0856. Available under: doi: 10.1016/j.jmr.2011.07.014

Zusammenfassung

Spin-label electron paramagnetic resonance (SL-EPR) spectroscopy has become a powerful and useful tool for studying structure and dynamics of biomacromolecules. However, utilizing these methods at physiological temperatures for in-cell studies is hampered by reduction of the nitroxide spin labels and thus short half-lives in the cellular environment. Consequently, reduction kinetics of two structurally different nitroxides was investigated in cell extracts of Xenopus laevis oocytes using rapid-scan cw-exper- iments at X-band. The five member heterocyclic ring nitroxide PCA (3-carboxy-2,2,5,5-tetramethylpyrro- lidinyl-1-oxy) under investigation features much higher stability against intracellular reduction than the six member ring analog TOAC (2,2,6,6-tetramethylpiperidine-N-oxyl-4-amino-4-carboxilic acid) and is therefore a suitable spin label type for in-cell EPR. The kinetic data can be described according to the Michaelis–Menten model and thus suggest an enzymatic or enzyme-mediated reduction process.

Fachgebiet (DDC)

570 Biowissenschaften, Biologie

Zitieren

ISO 690

AZARKH, Mykhailo, Oliver OKLE, Philipp EYRING, Daniel R. DIETRICH, Malte DRESCHER, 2011. Evaluation of spin labels for in-cell EPR by analysis of nitroxide reduction in cell extract of Xenopus laevis oocytes. In: Journal of Magnetic Resonance. 2011, 212(2), pp. 450-454. ISSN 1090-7807. eISSN 1096-0856. Available under: doi: 10.1016/j.jmr.2011.07.014

BibTex

@article{Azarkh2011-10Evalu-18165,
  year={2011},
  doi={10.1016/j.jmr.2011.07.014},
  title={Evaluation of spin labels for in-cell EPR by analysis of nitroxide reduction in cell extract of Xenopus laevis oocytes},
  number={2},
  volume={212},
  issn={1090-7807},
  journal={Journal of Magnetic Resonance},
  pages={450--454},
  author={Azarkh, Mykhailo and Okle, Oliver and Eyring, Philipp and Dietrich, Daniel R. and Drescher, Malte}
}

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