gem-Diethyl Pyrroline Nitroxide Spin Labels: Synthesis, EPR Characterization, Rotamer Libraries and Biocompatibility

Stephanie Bleicken, Tufa E. Assafa, Hui Zhang, Christina Elsner, Irina Ritsch, Maren Pink, Suchada Rajca, Gunnar Jeschke, Andrzej Rajca, Enrica Bordignon

Research output: Contribution to journalArticle

Abstract

The availability of bioresistant spin labels is crucial for the optimization of site-directed spin labeling protocols for EPR structural studies of biomolecules in a cellular context. As labeling can affect proteins’ fold and/or function, having the possibility to choose between different spin labels will increase the probability to produce spin-labeled functional proteins. Here, we report the synthesis and characterization of iodoacetamide- and maleimide-functionalized spin labels based on the gem-diethyl pyrroline structure. The two nitroxide labels are compared to conventional gem-dimethyl analogs by site-directed spin labeling (SDSL) electron paramagnetic resonance (EPR) spectroscopy, using two water soluble proteins: T4 lysozyme and Bid. To foster their use for structural studies, we also present rotamer libraries for these labels, compatible with the MMM software. Finally, we investigate the “true” biocompatibility of the gem-diethyl probes comparing the resistance towards chemical reduction of the NO group in ascorbate solutions and E. coli cytosol at different spin concentrations.

Original languageEnglish (US)
JournalChemistryOpen
DOIs
StatePublished - Jan 1 2019

Fingerprint

Gems
Spin Labels
Biocompatibility
Labeling
Paramagnetic resonance
Labels
Iodoacetamide
Proteins
Chemical resistance
Biomolecules
Muramidase
Escherichia coli
Availability
Spectroscopy
Water
pyrroline

Keywords

  • EPR
  • biocompatibility
  • nitroxide
  • pyrrolines
  • spin labeling

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

gem-Diethyl Pyrroline Nitroxide Spin Labels : Synthesis, EPR Characterization, Rotamer Libraries and Biocompatibility. / Bleicken, Stephanie; Assafa, Tufa E.; Zhang, Hui; Elsner, Christina; Ritsch, Irina; Pink, Maren; Rajca, Suchada; Jeschke, Gunnar; Rajca, Andrzej; Bordignon, Enrica.

In: ChemistryOpen, 01.01.2019.

Research output: Contribution to journalArticle

Bleicken, Stephanie ; Assafa, Tufa E. ; Zhang, Hui ; Elsner, Christina ; Ritsch, Irina ; Pink, Maren ; Rajca, Suchada ; Jeschke, Gunnar ; Rajca, Andrzej ; Bordignon, Enrica. / gem-Diethyl Pyrroline Nitroxide Spin Labels : Synthesis, EPR Characterization, Rotamer Libraries and Biocompatibility. In: ChemistryOpen. 2019.
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