Intermolecular dynamics studied by paramagnetic tagging |
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Authors: | Xingfu Xu Peter H. J. Keizers Wolfgang Reinle Frank Hannemann Rita Bernhardt Marcellus Ubbink |
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Affiliation: | 1.Institute of Chemistry,Leiden University,Leiden,The Netherlands;2.Naturwissenschaftlich-Technische Fakult?t III, Institut für Biochemie,Universit?t des Saarlandes,Saarbrücken,Germany |
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Abstract: | Yeast cytochrome c and bovine adrenodoxin form a dynamic electron transfer complex, which is a pure encounter complex. It is demonstrated that the dynamic nature of the interaction can readily be probed by using a rigid lanthanide tag attached to cytochrome c. The tag, Caged Lanthanide NMR Probe 5, induces pseudocontact shifts and residual dipolar couplings and does not perturb the binding interface. Due to the dynamics in the complex, residual dipolar couplings in adrenodoxin are very small. Simulation shows that cytochrome c needs to sample a large part of the surface of adrenodoxin to explain the small degree of alignment observed for adrenodoxin. The applied method provides a simple and straightforward way to observe dynamics in protein complexes or domain–domain mobility without the need for external alignment media. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. |
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Keywords: | Dynamics Lanthanide tag Encounter complex Residual dipolar couplings Paramagnetic NMR |
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