PT  - JOURNAL ARTICLE
AU  - Ralph Krafczyk
AU  - Jakub Macošek
AU  - Daniel Gast
AU  - Swetlana Wunder
AU  - Pravin Kumar Ankush Jagtap
AU  - Prithiba Mitra
AU  - Amit Kumar Jha
AU  - Jüergen Rohr
AU  - Anja Hoffmann-Röeder
AU  - Kirsten Jung
AU  - Janosch Hennig
AU  - Jüergen Lassak
TI  - Structural basis for EarP-mediated arginine glycosylation of translation elongation factor EF-P
AID  - 10.1101/116038
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 116038
4099  - http://biorxiv.org/content/early/2017/03/11/116038.short
4100  - http://biorxiv.org/content/early/2017/03/11/116038.full
AB  - Glycosylation is a universal strategy to post-translationally modify proteins. The recently discovered arginine rhamnosylation activates the polyproline specific bacterial translation elongation factor EF-P. EF-P is rhamnosylated on arginine 32 by the glycosyltransferase EarP. However, the enzymatic mechanism remains elusive. In the present study, we solved the crystal structure of EarP from Pseudomonas putida. The enzyme is composed of two opposing domains with Rossmann-folds, thus constituting a GT-B glycosyltransferase. While TDP-rhamnose is located within a highly conserved pocket of the C-domain, EarP recognizes the EF-P via its KOW-like N-domain. Based on our structural data combined with an in vitro /in vivo enzyme characterization, we propose a mechanism of inverting arginine glycosylation. As EarP is essential for pathogenicity in P. aeruginosa our study provides the basis for targeted inhibitor design.