Genetic recoding to dissect the roles of site-specific protein O-GlcNAcylation

Andrii Gorelik, Sergio Galan Bartual, Vladimir Borodkin, Joby Varghese, Andrew Ferenbach, Daan van Aalten (Lead / Corresponding author)

Research output: Contribution to journalArticle

4 Citations (Scopus)
33 Downloads (Pure)

Abstract

Modification of specific Ser and Thr residues of nucleocytoplasmic proteins with O-GlcNAc, catalyzed by O-GlcNAc transferase (OGT), is an abundant posttranslational event essential for proper animal development and is dysregulated in various diseases. Due to the rapid concurrent removal by the single O-GlcNAcase (OGA), precise functional dissection of site-specific O-GlcNAc modification in vivo is currently not possible without affecting the entire O-GlcNAc proteome. Exploiting the fortuitous promiscuity of OGT, we show that S-GlcNAc is a hydrolytically stable and accurate structural mimic of O-GlcNAc that can be encoded in mammalian systems with CRISPR-Cas9 in an otherwise unperturbed O-GlcNAcome. Using this approach, we target an elusive Ser 405 O-GlcNAc site on OGA, showing that this site-specific modification affects OGA stability.

Original languageEnglish
Pages (from-to)1071-1077
Number of pages7
JournalNature Structural & Molecular Biology
Volume26
Issue number11
Early online date6 Nov 2019
DOIs
Publication statusPublished - Nov 2019

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