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Scientific article
English

Distance determination between low-spin ferric haem and nitroxide spin label using DEER: the neuroglobin case

Published inMolecular physics, vol. 111, no. 18-19, p. 2855-2864
Publication date2013-07-08
First online date2013-07-08
Abstract

This work demonstrates for the first time the feasibility of using double electron–electron resonance (DEER) to determine the inter-spin distance between nitroxide spin labels and low-spin (S = 1/2) ferric haem centres. For these means, two human neuroglobin variants were spin labelled leading to singly labelled haem proteins with the nitroxide label on one of the natural Cys residues (Cys55 or Cys120). Room-temperature electron paramagnetic resonance was used to characterise the mobility of the nitroxide labels and X- and Q-band DEER experiments were performed to detect nitroxide–haem distances. Effects of residual nuclear modulations in the DEER traces were carefully evaluated. The DEER-derived distances were compared with theoretical predictions from an X-ray diffraction structure of human neuroglobin using a rotamer library approach as well as with distance information obtained from electron relaxation measurements. The structural biological implications of the spin-labelled side chains’ dynamics and of the obtained distances are also discussed.

eng
Keywords
  • EPR
  • Distance mesurements
  • Neuroglobin
Affiliation Not a UNIGE publication
Citation (ISO format)
EZHEVSKAYA, M. et al. Distance determination between low-spin ferric haem and nitroxide spin label using DEER: the neuroglobin case. In: Molecular physics, 2013, vol. 111, n° 18-19, p. 2855–2864. doi: 10.1080/00268976.2013.813592
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ISSN of the journal0026-8976
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