期刊
NATURE METHODS
卷 6, 期 7, 页码 532-U94出版社
NATURE PUBLISHING GROUP
DOI: 10.1038/nmeth.1341
关键词
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资金
- Howard Hughes Medical Institute
- National Institutes of Health (NIH) [EY10329, R01 NS038631, 1K99NS064213]
- National Eye Institute
- Jane Coffin Childs Foundation
Visualizing conformational dynamics in proteins has been difficult, and the atomic-scale motions responsible for the behavior of most allosteric proteins are unknown. Here we report that fluorescence resonance energy transfer (FRET) between a small fluorescent dye and a nickel ion bound to a dihistidine motif can be used to monitor small structural rearrangements in proteins. This method provides several key advantages over classical FRET, including the ability to measure the dynamics of close-range interactions, the use of small probes with short linkers, a low orientation dependence, and the ability to add and remove unique tunable acceptors. We used this 'transition metal ion FRET' approach along with X-ray crystallography to determine the structural changes of the gating ring of the mouse hyperpolarization-activated cyclic nucleotide-regulated ion channel HCN2. Our results suggest a general model for the conformational switch in the cyclic nucleotide-binding site of cyclic nucleotide-regulated ion channels.
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