caa a22 4500 445325062 CHVBK 20180317142717.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s10858-011-9471-y doi (NATIONALLICENCE)springer-10.1007/s10858-011-9471-y Integrated analysis of the conformation of a protein-linked spin label by crystallography, EPR and NMR spectroscopy [Elektronische Daten] [Tim Gruene, Min-Kyu Cho, Irina Karyagina, Hai-Young Kim, Christian Grosse, Karin Giller, Markus Zweckstetter, Stefan Becker] 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 protein-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. The Author(s), 2011 Spin label nationallicence Crystal structure nationallicence MTSL nationallicence Protein nationallicence NMR nationallicence MTSL : (1-oxy-2,2,5,5-tetramethyl-D-pyrroline-3-methyl)-methanethiosulfonate nationallicence HSQC : Heteronuclear single quantum coherence nationallicence EPR : Electron paramagnetic resonance nationallicence DEER : Double electron electron resonance nationallicence PRE : Paramagnetic relaxation enhancement nationallicence Gruene Tim Department of Structural Chemistry, University of Göttingen, Tammannstraße 4, 37077, Göttingen, Germany aut Cho Min-Kyu Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut Karyagina Irina Max Planck Institute for Biophysical Chemistry, AG Electron Spin Resonance Spectroscopy, Am Faßberg 11, 37077, Göttingen, Germany aut Kim Hai-Young Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut Grosse Christian Department of Structural Chemistry, University of Göttingen, Tammannstraße 4, 37077, Göttingen, Germany aut Giller Karin Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut Zweckstetter Markus Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut Becker Stefan Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut Journal of Biomolecular NMR Springer Netherlands 49/2(2011-02-01), 111-119 0925-2738 49:2<111 2011 49 10858 https://doi.org/10.1007/s10858-011-9471-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10858-011-9471-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Gruene Tim Department of Structural Chemistry, University of Göttingen, Tammannstraße 4, 37077, Göttingen, Germany aut NATIONALLICENCE 700 1- Cho Min-Kyu Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut NATIONALLICENCE 700 1- Karyagina Irina Max Planck Institute for Biophysical Chemistry, AG Electron Spin Resonance Spectroscopy, Am Faßberg 11, 37077, Göttingen, Germany aut NATIONALLICENCE 700 1- Kim Hai-Young Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut NATIONALLICENCE 700 1- Grosse Christian Department of Structural Chemistry, University of Göttingen, Tammannstraße 4, 37077, Göttingen, Germany aut NATIONALLICENCE 700 1- Giller Karin Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut NATIONALLICENCE 700 1- Zweckstetter Markus Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut NATIONALLICENCE 700 1- Becker Stefan Department of NMR based Structural Biology, Max Planck Institute for Biophysical Chemistry, Am Faßberg 11, 37077, Göttingen, Germany aut NATIONALLICENCE 773 0- Journal of Biomolecular NMR Springer Netherlands 49/2(2011-02-01), 111-119 0925-2738 49:2<111 2011 49 10858 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer