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Efficient localization of a native metal ion within a protein by Cu2+-based EPR distance measurements

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Abstract

Electron paramagnetic resonance (EPR) based distance measurements have been exploited to measure protein–protein docking, protein–DNA interactions, substrate binding and metal coordination sites. Here, we use EPR to locate a native paramagnetic metal binding site in a protein with less than 2 Å resolution. We employ a rigid Cu2+ binding motif, the double histidine (dHis) motif, in conjunction with double electron electron resonance (DEER) spectroscopy. Specifically, we utilize a multilateration approach to elucidate the native Cu2+ binding site in the immunoglobulin binding domain of protein G. Notably, multilateration performed with the dHis motif required only the minimum number of four distance constraints, whereas comparable studies using flexible nitroxide-based spin labels require many more for similar precision. This methodology demonstrates a significant increase in the efficiency of structural determinations via EPR distance measurements using the dHis motif.

Graphical abstract: Efficient localization of a native metal ion within a protein by Cu2+-based EPR distance measurements

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Publication details

The article was received on 19 Nov 2018, accepted on 01 Feb 2019 and first published on 04 Feb 2019


Article type: Paper
DOI: 10.1039/C8CP07143H
Citation: Phys. Chem. Chem. Phys., 2019, Advance Article

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    Efficient localization of a native metal ion within a protein by Cu2+-based EPR distance measurements

    A. Gamble Jarvi, T. F. Cunningham and S. Saxena, Phys. Chem. Chem. Phys., 2019, Advance Article , DOI: 10.1039/C8CP07143H

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