Gadolinium tagging for high-precision measurements of 6 nm distances in protein assemblies by EPR
Date
2011
Authors
Yagi, Hiromasa
Banerjee, Debamalya
Graham, Bim
Goldfarb, Daniella
Otting, Gottfried
Huber, Thomas
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American Chemical Society
Abstract
Double electron-electron resonance (DEER) distance measurements of a protein complex tagged with two Gd3+ chelates developed for rigid positioning of the metal ion are shown to deliver outstandingly accurate distance measurements in the 6 nm range. The accuracy was assessed by comparison with modeled distance distributions based on the three-dimensional molecular structures of the protein and the tag and further comparison with paramagnetic NMR data. The close agreement between the predicted and experimentally measured distances opens new possibilities for investigating the structure of biomolecular assemblies. As an example, we show that the dimer interface of rat ERp29 in solution is the same as that determined previously for human ERp29 in the single crystal.
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Keywords: Bio-molecular; Dimer interface; Distance distributions; Double electron-electron resonance; High-precision measurement; Paramagnetic NMR; Protein assembly; Protein complexes; Dimers; Electron resonance; Electron spin resonance spectroscopy; Gadolinium; Me
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Journal of the American Chemical Society
Type
Journal article
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2037-12-31
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