19 F Paramagnetic Relaxation-Based NMR for Quaternary Structural Restraints of Ion Channels.
ACS chemical biology(2019)
摘要
Quaternary distance restraints are essential to define the three-dimensional structures of protein assemblies. These distances often fall within a range of 10-18 Å, which challenges the high and low measurement limits of conventional nuclear magnetic resonance (NMR) spectroscopy and double electron-electron resonance electron spin resonance spectroscopy. Here, we report the use of F paramagnetic relaxation enhancement (PRE) NMR in combination with F/paramagnetic labeling to equivalent sites in different subunits of a protein complex in micelles to determine inter-subunit distances. Feasibility of this strategy was evaluated on a pentameric ligand-gated ion channel, for which we found excellent agreement of the F PRE NMR results with previous structural information. The study suggests that F PRE NMR is a viable tool in extracting distance restraints to define quaternary structures.
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关键词
nmr,quaternary structural restraints,relaxation-based
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