Journal
JOURNAL OF BIOMOLECULAR NMR
Volume 43, Issue 4, Pages 247-254Publisher
SPRINGER
DOI: 10.1007/s10858-009-9308-0
Keywords
Dynamics; Lanthanide tag; Encounter complex; Residual dipolar couplings; Paramagnetic NMR
Categories
Funding
- Fonds der chem. Industrie
- Volkswagenstiftung [I/80854]
- Netherlands Organisation for Scientific Research [700.50.026, 700.52.425]
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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.
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