Title data
Li, Xiang ; Krafczyk, Ralph ; Macošek, Jakub ; Li, Yu-Lei ; Zou, Yan ; Simon, Bernd ; Pan, Xing ; Wu, Qiu-Ye ; Yan, Fang ; Li, Shan ; Hennig, Janosch ; Jung, Kirsten ; Lassak, Jürgen ; Hu, Hong-Gang:
Resolving the α-glycosidic linkage of arginine-rhamnosylated translation elongation factor P triggers generation of the first Argᴿʰᵃ specific antibody.
In: Chemical Science.
Vol. 7
(2016)
Issue 12
.
- pp. 6995-7001.
ISSN 2041-6539
DOI: https://doi.org/10.1039/c6sc02889f
Abstract in another language
A previously discovered posttranslational modification strategy - arginine rhamnosylation - is essential for elongation factor P (EF-P) dependent rescue of polyproline stalled ribosomes in clinically relevant species such as Pseudomonas aeruginosa and Neisseria meningitidis. However, almost nothing is known about this new type of N-linked glycosylation. In the present study we used NMR spectroscopy to show for the first time that the α anomer of rhamnose is attached to Arg32 of EF-P, demonstrating that the corresponding glycosyltransferase EarP inverts the sugar of its cognate substrate dTDP-β-l-rhamnose. Based on this finding we describe the synthesis of an α-rhamnosylated arginine containing peptide antigen in order to raise the first anti-rhamnosyl arginine specific antibody (anti-ArgRha). Using ELISA and Western Blot analyses we demonstrated both its high affinity and specificity without any cross-reactivity to other N-glycosylated proteins. Having the anti-ArgRha at hand we were able to visualize endogenously produced rhamnosylated EF-P. Thus, we expect the antibody to be not only important to monitor EF-P rhamnosylation in diverse bacteria but also to identify further rhamnosyl arginine containing proteins. As EF-P rhamnosylation is essential for pathogenicity, our antibody might also be a powerful tool in drug discovery.
Further data
Item Type: | Article in a journal |
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Refereed: | Yes |
Institutions of the University: | Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry with an Emphasis on Biophysical Chemistry > Chair Biochemistry with an Emphasis on Biophysical Chemistry - Univ.-Prof. Dr. Janosch Hennig Faculties Faculties > Faculty of Biology, Chemistry and Earth Sciences Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry Faculties > Faculty of Biology, Chemistry and Earth Sciences > Department of Chemistry > Chair Biochemistry with an Emphasis on Biophysical Chemistry |
Result of work at the UBT: | No |
DDC Subjects: | 500 Science > 540 Chemistry 500 Science > 570 Life sciences, biology |
Date Deposited: | 07 Oct 2021 12:26 |
Last Modified: | 07 Oct 2021 12:26 |
URI: | https://eref.uni-bayreuth.de/id/eprint/67233 |