Characterization of the Walker A Motif of MsbA Using Site-Directed Spin Labeling Electron Paramagnetic Resonance Spectroscopy†

Biochemistry ◽  
2005 ◽  
Vol 44 (14) ◽  
pp. 5503-5509 ◽  
Author(s):  
Adam H. Buchaklian ◽  
Candice S. Klug
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Electron Paramagnetic Resonance Spectroscopy ◽  
Spin Labeling ◽  
Resonance Spectroscopy ◽  
Paramagnetic Resonance ◽  
Site Directed Spin Labeling ◽  
Electron Paramagnetic

scholarly journals Site Selective and Efficient Spin Labeling of Proteins with a Maleimide-Functionalized Trityl Radical for Pulsed Dipolar EPR Spectroscopy

Molecules ◽  
2019 ◽  
Vol 24 (15) ◽  
pp. 2735 ◽  
Author(s):  
J. Jacques Jassoy ◽  
Caspar A. Heubach ◽  
Tobias Hett ◽  
Frédéric Bernhard ◽  
Florian R. Haege ◽  
...  
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Electron Paramagnetic Resonance Spectroscopy ◽  
Spin Labeling ◽  
Single Frequency ◽  
Spin Labels ◽  
Resonance Spectroscopy ◽  
Paramagnetic Resonance ◽  
Site Directed Spin Labeling ◽  
Site Selective ◽  
Electron Paramagnetic

Pulsed dipolar electron paramagnetic resonance spectroscopy (PDS) in combination with site-directed spin labeling (SDSL) of proteins and oligonucleotides is a powerful tool in structural biology. Instead of using the commonly employed gem-dimethyl-nitroxide labels, triarylmethyl (trityl) spin labels enable such studies at room temperature, within the cells and with single-frequency electron paramagnetic resonance (EPR) experiments. However, it has been repeatedly reported that labeling of proteins with trityl radicals led to low labeling efficiencies, unspecific labeling and label aggregation. Therefore, this work introduces the synthesis and characterization of a maleimide-functionalized trityl spin label and its corresponding labeling protocol for cysteine residues in proteins. The label is highly cysteine-selective, provides high labeling efficiencies and outperforms the previously employed methanethiosulfonate-functionalized trityl label. Finally, the new label is successfully tested in PDS measurements on a set of doubly labeled Yersinia outer protein O (YopO) mutants.

Localization of metal ions in biomolecules by means of pulsed dipolar EPR spectroscopy

2021 ◽  
Author(s):  
Dinar Abdullin ◽  
Olav Schiemann
Keyword(s):  
Electron Paramagnetic Resonance ◽  
Metal Ions ◽  
Epr Spectroscopy ◽  
Electron Paramagnetic Resonance Spectroscopy ◽  
Spin Labeling ◽  
Resonance Spectroscopy ◽  
Paramagnetic Resonance ◽  
Paramagnetic Metal ◽  
Site Directed Spin Labeling ◽  
Electron Paramagnetic

A method is introduced in which paramagnetic metal ions are localized by means of trilateration using a combination of site-directed spin labeling and pulsed dipolar electron paramagnetic resonance spectroscopy.

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