Spin labelling via metabolic glycoengineering for studying post-translational protein modification by electron paramagnetic resonance spectroscopy

13 May 2022, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

Post-translational modifications are involved in many cellular processes. The addition of O-linked β-N-acetylglucosamine (O-GlcNAc) is a ubiquitous post-translational modification and essential for regulatory processes in mammalian cells. Here, we demonstrate specific spin labelling of post-translational modifications, namely glycosylation modifications, via metabolic glycoengineering and subsequent copper-catalyzed azide-alkyne cycloaddition. Electron paramagnetic resonance spectroscopy and subsequent quantitative spectral simulations allow for detection of the labelled post-translational modifications.

Keywords

Post-translational modifications
O-GlcNAcylation
Metabolic glycoengineering (MGE)
copper-catalyzed azide-alkyne cycloaddition (CuAAC)
spin labelling
electron paramagnetic resonance (EPR) spectroscopy

Supplementary materials

Title
Description
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Title
Spin labelling via metabolic glycoengineering for studying post-translational protein modification by electron paramagnetic resonance spectroscopy
Description
Supporting information includes the details of the experimental procedure.
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