Combining site-directed spin labeling in vivo and in-cell EPR distance determination

Widder, Pia; Schuck, Julian; Summerer, Daniel; Drescher, Malte

Combining site-directed spin labeling in vivo and in-cell EPR distance determination

Dateien

Widder_2-1fsurzavjju712.pdfGröße: 2.18 MBDownloads: 283

Datum

2020

Open Access-Veröffentlichung

Open Access Hybrid

Sammlungen

Chemie

Publikationstyp

Zeitschriftenartikel

Publikationsstatus

Published

Erschienen in

Physical Chemistry, Chemical Physics (PCCP). Royal Society of Chemistry (RSC). 2020, 22(9), pp. 4875-4879. ISSN 1463-9076. eISSN 1463-9084. Available under: doi: 10.1039/c9cp05584c

Zusammenfassung

Structural studies on proteins directly in their native environment are required for a comprehensive understanding of their function. Electron paramagnetic resonance (EPR) spectroscopy and in particular double electron-electron resonance (DEER) distance determination are suited to investigate spin-labeled proteins directly in the cell. The combination of intracellular bioorthogonal labeling with in-cell DEER measurements does not require additional purification or delivery steps of spin-labeled protein to the cells. In this study, we express eGFP in E. coli and use copper-catalyzed azide-alkyne cycloaddition (CuAAC) for the site-directed spin labeling of the protein in vivo, followed by in-cell EPR distance determination. Inter-spin distance measurements of spin-labeled eGFP agree with in vitro measurements and calculations based on the rotamer library of the spin label.

Fachgebiet (DDC)

540 Chemie

Zitieren

ISO 690

WIDDER, Pia, Julian SCHUCK, Daniel SUMMERER, Malte DRESCHER, 2020. Combining site-directed spin labeling in vivo and in-cell EPR distance determination. In: Physical Chemistry, Chemical Physics (PCCP). Royal Society of Chemistry (RSC). 2020, 22(9), pp. 4875-4879. ISSN 1463-9076. eISSN 1463-9084. Available under: doi: 10.1039/c9cp05584c

BibTex

@article{Widder2020-03-07Combi-49228,
  year={2020},
  doi={10.1039/c9cp05584c},
  title={Combining site-directed spin labeling in vivo and in-cell EPR distance determination},
  number={9},
  volume={22},
  issn={1463-9076},
  journal={Physical Chemistry, Chemical Physics (PCCP)},
  pages={4875--4879},
  author={Widder, Pia and Schuck, Julian and Summerer, Daniel and Drescher, Malte}
}

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