Isoindoline‐Based Nitroxides as Bioresistant Spin Labels for Protein Labeling through Cysteines and Alkyne‐Bearing Noncanonical Amino Acids | Zendy

Braun Theresa Sophie | Zendy; Widder Pia | Zendy; Osswald Uwe | Zendy; Groß Lina | Zendy; Williams Lara | Zendy; Schmidt Moritz | Zendy; Helmle Irina | Zendy; Summerer Daniel | Zendy; Drescher Malte | Zendy

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Isoindoline‐Based Nitroxides as Bioresistant Spin Labels for Protein Labeling through Cysteines and Alkyne‐Bearing Noncanonical Amino Acids

Author(s) -

Braun Theresa Sophie,

Widder Pia,

Osswald Uwe,

Groß Lina,

Williams Lara,

Schmidt Moritz,

Helmle Irina,

Summerer Daniel,

Drescher Malte

Publication year - 2020

Publication title -

chembiochem

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.05

H-Index - 126

eISSN - 1439-7633

pISSN - 1439-4227

DOI - 10.1002/cbic.201900537

Subject(s) - site directed spin labeling , chemistry , electron paramagnetic resonance , isoindoline , nitroxide mediated radical polymerization , azide , spin label , bioorthogonal chemistry , cysteine , conformational isomerism , combinatorial chemistry , stereochemistry , biochemistry , click chemistry , nuclear magnetic resonance , organic chemistry , molecule , polymerization , enzyme , physics , radical polymerization , membrane , polymer

Electron paramagnetic resonance (EPR) spectroscopy in combination with site‐directed spin labeling (SDSL) is a powerful tool in protein structural research. Nitroxides are highly suitable spin labeling reagents, but suffer from limited stability, particularly in the cellular environment. Herein we present the synthesis of a maleimide‐ and an azide‐modified tetraethyl‐shielded isoindoline‐based nitroxide (M‐ and Az‐TEIO) for labeling of cysteines or the noncanonical amino acid para ‐ethynyl‐ l ‐phenylalanine ( p ENF). We demonstrate the high stability of TEIO site‐specifically attached to the protein thioredoxin (TRX) against reduction in prokaryotic and eukaryotic environments, and conduct double electron–electron resonance (DEER) measurements. We further generate a rotamer library for the new residue p ENF‐Az‐TEIO that affords a distance distribution that is in agreement with the measured distribution.

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