Probing the Iron−Substrate Orientation for Taurine/α-Ketoglutarate Dioxygenase Using Deuterium Electron Spin Echo Envelope Modulation Spectroscopy
- 1 May 2007
- journal article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 46 (20), 5951-5959
- https://doi.org/10.1021/bi700562t
Abstract
The structural relationship between substrate taurine and the non-heme Fe(II) center of taurine/α-ketoglutarate (αKG) dioxygenase (TauD) was measured using electron spin echo envelope modulation (ESEEM) spectroscopy. Studies were conducted on TauD samples treated with NO, cosubstrate αKG, and either protonated or specifically deuterated taurine. Stimulated echo ESEEM data were divided to eliminate interference from 1H and 14N modulations and accentuate modulations from 2H. For taurine that was deuterated at the C1 position (adjacent to the sulfonate group), 2H ESEEM spectra show features that arise from dipole−dipole and deuterium nuclear quadrupole interactions from a single deuteron. Parallel measurements taken for taurine deuterated at both C1 and C2 show an additional ESEEM feature at the deuterium Larmor frequency. Analysis of these data at field positions ranging from g = 4 to g = 2 have allowed us to define the orientation of substrate taurine with respect to the magnetic axes of the Fe(II)−NO, S = 3/2, paramagnetic center. These results are discussed in terms of previous X-ray crystallographic studies and the proposed catalytic mechanism for this family of enzymes.Keywords
This publication has 1 reference indexed in Scilit:
- Principles of Pulse Electron Paramagnetic ResonancePublished by Oxford University Press (OUP) ,2001