Glycosylation is a universal strategy to posttranslationally 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 B pattern-type glycosyltransferase (GT-B). While dTDP-β-L-rhamnose is located within a highly conserved pocket of the C-domain, EarP recognizes the KOW-like N-domain of EF-P. Based on our data, we propose a structural model for arginine glycosylation by EarP. As EarP is essential for pathogenicity in P. aeruginosa, our study provides the basis for targeted inhibitor design.

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Published in mBio, v. 8, issue 5, e01412-17, p. 1-20.

Copyright © 2017 Krafczyk et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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J.L., K.J., and A.H.R. gratefully acknowledge financial support from the DFG Research Training Group GRK2062 (Molecular Principles of Synthetic Biology). Moreover, J.L. is grateful for DFG grant LA 3658/1-1. J.H. acknowledges support from the European Molecular Biology Laboratory (EMBL). P.K.A.J. acknowledges EMBL and the EU Marie Curie Actions Cofund for an EIPOD fellowship. K.J. and A.H.R. additionally thank the Center for integrated Protein Science Munich (Cluster of Excellence grant Exc114/2). The work of J.R., P.M., and A.K.J. was supported by U.S. National Institutes of Health grant GM 105977.

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Atomic coordinates and structure factors for the reported crystal structures have been deposited with the Protein Data Bank under accession number 5NV8.

Supplemental material for this article may be found at https://doi.org/10.1128/mBio .01412-17.