Abstract Long-range structural information derived from paramagnetic relaxation enhancement observed in the presence of a paramagnetic nitroxide radical is highly useful for structural characterization of globular, modular and intrinsically disordered proteins, as well as proteinâ??protein and protein-DNA complexes. Here we characterized the conformation of a spin-label attached to the homodimeric protein CylR2 using a combination of X-ray crystallography, electron paramagnetic resonance (EPR) and NMR spectroscopy. Close agreement was found between the conformation of the spin label observed in the crystal structure with interspin distances measured by EPR and signal broadening in NMR spectra, suggesting that the conformation seen in the crystal structure is also preferred in solution. In contrast, conformations of the spin label observed in crystal structures of T4 lysozyme are not in agreement with the paramagnetic relaxation enhancement observed for spin-labeled CylR2 in solution. Our data demonstrate that accurate positioning of the paramagnetic center is essential for high-resolution structure determination.
Content Type Journal Article
Pages 1-9
DOI 10.1007/s10858-011-9471-y
Authors
Tim Gruene, Department of Structural Chemistry, University of Göttingen, Tammannstra�e 4, 37077 Göttingen, Germany
Min-Kyu Cho, Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fa�berg 11, 37077 Göttingen, Germany
Irina Karyagina, Max Planck Institute for Biophysical Chemistry, AG Electron Spin Resonance Spectroscopy, Am Fa�berg 11, 37077 Göttingen, Germany
Hai-Young Kim, Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fa�berg 11, 37077 Göttingen, Germany
Christian Grosse, Department of Structural Chemistry, University of Göttingen, Tammannstra�e 4, 37077 Göttingen, Germany
Karin Giller, Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fa�berg 11, 37077 Göttingen, Germany
Markus Zweckstetter, Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fa�berg 11, 37077 Göttingen, Germany
Stefan Becker, Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Fa�berg 11, 37077 Göttingen, Germany
Orientation Selective DEER Measurements on Vinculin Tail at X-Band Frequencies Reveal Spin Label Orientations
Orientation Selective DEER Measurements on Vinculin Tail at X-Band Frequencies Reveal Spin Label Orientations
Publication year: 2012
Source: Journal of Magnetic Resonance, Available online 8 January 2012</br>
Christoph*Abé, Daniel*Klose, Franziska*Dietrich, Wolfgang H.*Ziegler, Yevhen*Polyhach, ...</br>
Double electron electron resonance (DEER) spectroscopy has been established as a valuable method to determine distances between spin labels bound to protein molecules. Caused by selective excitation of molecular orientations DEER primary data also depend on the mutual orientation of...
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Combination of NMR spectroscopy and X-ray crystallography offers unique advantages for elucidation of the structural basis of protein complex assembly.
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Integrated analysis of the conformation of a protein-linked spin label by crystallography, EPR and NMR spectroscopy.
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Authors: Gruene T, Cho MK, Karyagina I, Kim HY, Grosse C, Giller K, Zweckstetter M, Becker S
Long-range structural information derived from paramagnetic relaxation enhancement observed in the presence of a paramagnetic nitroxide radical is highly useful for structural characterization of...
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Integrated analysis of the conformation of a protein-linked spin label by crystallography, EPR and NMR spectroscopy
Linear Mode
