Intermolecular dynamics studied by paramagnetic tagging.
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.
Xu
Xingfu
X
Institute of Chemistry, Leiden University, P.O. Box 9502, 2300 RA, Leiden, The Netherlands.
Keizers
Peter H J
PH
Reinle
Wolfgang
W
Hannemann
Frank
F
Bernhardt
Rita
R
Ubbink
Marcellus
M
D000325:Adrenodoxin / Q000737:chemistry* / Q000378:metabolism*; D000818:Animals; D002417:Cattle; D045304:Cytochromes c / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000378:metabolism; D028581:Lanthanoid Series Elements / Q000378:metabolism*; D008956:Models, Chemical; D016297:Mutagenesis, Site-Directed; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000378:metabolism
D005656:Fungal Proteins; D028581:Lanthanoid Series Elements; D009587:Nitrogen Isotopes; D011506:Proteins; D000325:Adrenodoxin; D045304:Cytochromes c