Gadolinium Tagging for High-Precision Measurements of 6 nm Distances in Protein Assemblies by EPR

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• 作者：Hiromasa Yagi ; Debamalya Banerjee ; Bim Graham ; Thomas Huber ; Daniella Goldfarb ; Gottfried Otting
• 刊名：Journal of the American Chemical Society
• 出版年：2011
• 出版时间：July 13, 2011
• 年：2011
• 卷：133
• 期：27
• 页码：10418-10421
• 全文大小：769K
• 年卷期：v.133,no.27(July 13, 2011)
• ISSN：1520-5126

文摘

Double electron鈥揺lectron 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.