Title
Distance determination between low-spin ferric haem and nitroxide spin label using DEER : the neuroglobin case Distance determination between low-spin ferric haem and nitroxide spin label using DEER : the neuroglobin case
Author
Faculty/Department
Faculty of Sciences. Physics
Faculty of Pharmaceutical, Biomedical and Veterinary Sciences . Biomedical Sciences
Publication type
article
Publication
London ,
Subject
Physics
Chemistry
Source (journal)
Molecular physics. - London
Volume/pages
111(2013) :18-19 , p. 2855-2864
ISSN
0026-8976
0026-8976
ISI
000325389800019
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
This work demonstrates for the first time the feasibility of using double electronelectron 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 nitroxidehaem 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.
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