A Genetically Encoded Spin Label for Electron Paramagnetic Resonance Distance Measurements

2014 ◽  
Vol 136 (4) ◽  
pp. 1238-1241 ◽  
Author(s):  
Moritz J. Schmidt ◽  
Julia Borbas ◽  
Malte Drescher ◽  
Daniel Summerer
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Spin Label ◽  
Distance Measurements ◽  
Paramagnetic Resonance ◽  
Electron Paramagnetic

scholarly journals Pulsed ELDOR Measurement of the Distance Between a Spin-Label and Copper (II) Centre in the Copper Loaded R48C Mutant of N. gonorrhoeae Ferric Binding Protein

2018 ◽  
Author(s):  
Matt Bawn ◽  
Justin Bradley ◽  
Fraser MacMillan
Keyword(s):  
Spin Label ◽  
Binding Protein ◽  
Double Resonance ◽  
Distance Measurements ◽  
Paramagnetic Resonance ◽  
Distance Determination ◽  
Pulsed Epr ◽  
Ferric Binding Protein ◽  
Relaxation Experiments ◽  
Electron Paramagnetic

AbstractDistance determination in proteins and biomolecules using pulsed EPR (electron paramagnetic resonance) techniques is becoming an increasingly popular and accessible technique. PELDOR (pulsed electron-electron double resonance), is a technique designed for distance determination over a nanoscopic scale. Here, ferric binding protein (Fbp) is used to demonstrate the practicability of this technique to Cu (II) Metalloproteins. PELDOR is usually applied to bi-radicals or endogenous radicals, and distance determination using pulsed EPR of metal containing centres in biomolecules has been restricted to relaxation experiments. PELDOR distance measurements between a Cu (II) ion and a nitroxide have previously only been reported for model compounds [1, 2].Fbp as the name suggests usually, contains a Fe (III) ion centre. For the purposes of this investigation the Fe (III) ion was removed and replaced by a Cu (II) ion, after a nitroxide spin-label was added to the Fbp using of site directed spin-labelling (SDSL). PELDOR was then applied to measure the distance between the two centres.Simulation methods were then employed to fully investigate these data and allow a quantitative interpretation of the copper nitroxide PELDOR data. The observed PELDOR time traces were analysed using DEER analysis[3].

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