Development of ADPribosyl Ubiquitin Analogues to Study Enzymes Involved in Legionella Infection

Overview

Journal Chemistry

Specialty Chemistry

Date 2020 Oct 19

PMID 33075184

Citations 7

Authors

Robbert Q Kim

Mohit Misra

Alexis Gonzalez

Ines Tomaskovic

Donghyuk Shin

Hermann Schindelin

Dmitri V Filippov

Huib Ovaa

Ivan Dikic

Gerbrand J van der Heden van Noort

Affiliations

Soon will be listed here.

Abstract

Legionnaires' disease is caused by infection with the intracellularly replicating Gram-negative bacterium Legionella pneumophila. This pathogen uses an unconventional way of ubiquitinating host proteins by generating a phosphoribosyl linkage between substrate proteins and ubiquitin by making use of an ADPribosylated ubiquitin (Ub ) intermediate. The family of SidE effector enzymes that catalyze this reaction is counteracted by Legionella hydrolases, which are called Dups. This unusual ubiquitination process is important for Legionella proliferation and understanding these processes on a molecular level might prove invaluable in finding new treatments. Herein, a modular approach is used for the synthesis of triazole-linked Ub , and analogues thereof, and their affinity towards the hydrolase DupA is determined and hydrolysis rates are compared to natively linked Ub . The inhibitory effects of modified Ub on the canonical eukaryotic E1-enzyme Uba1 are investigated and rationalized in the context of a high-resolution crystal structure reported herein. Finally, it is shown that synthetic Ub analogues can be used to effectively pull-down overexpressed DupA from cell lysate.

Citing Articles

Covalent Probes To Capture Dup Effector Enzymes.

Kloet M, Mukhopadhyay R, Mukherjee R, Misra M, Jeong M, Talavera Ormeno C J Am Chem Soc. 2024; 146(39):26957-26964.

PMID: 39288007 PMC: 11450808. DOI: 10.1021/jacs.4c08168.

Capturing effector enzymes using a ubiquitin derived photo-activatable probe.

Kloet M, van der Heden van Noort G Front Mol Biosci. 2024; 11:1422034.

PMID: 39044841 PMC: 11263097. DOI: 10.3389/fmolb.2024.1422034.

Arginine ADP-Ribosylation: Chemical Synthesis of Post-Translationally Modified Ubiquitin Proteins.

Voorneveld J, Kloet M, Wijngaarden S, Kim R, Moutsiopoulou A, Verdegaal M J Am Chem Soc. 2022; 144(45):20582-20589.

PMID: 36318515 PMC: 9673145. DOI: 10.1021/jacs.2c06249.

A new dawn beyond lysine ubiquitination.

Squair D, Virdee S Nat Chem Biol. 2022; 18(8):802-811.

PMID: 35896829 DOI: 10.1038/s41589-022-01088-2.

Mimetics of ADP-Ribosylated Histidine through Copper(I)-Catalyzed Click Chemistry.

Minnee H, Rack J, van der Marel G, Overkleeft H, Codee J, Ahel I Org Lett. 2022; 24(21):3776-3780.

PMID: 35587229 PMC: 9171823. DOI: 10.1021/acs.orglett.2c01300.

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