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Publication Abstract from PubMed

Upon infection of host cells, Legionella pneumophila releases a multitude of effector enzymes into the cell's cytoplasm that hijack a plethora of cellular activities, including the host ubiquitination pathways. Effectors belonging to the SidE-family are involved in noncanonical serine phosphoribosyl ubiquitination of host substrate proteins contributing to the formation of a Legionella-containing vacuole that is crucial in the onset of Legionnaires' disease. This dynamic process is reversed by effectors called Dups that hydrolyze the phosphodiester in the phosphoribosyl ubiquitinated protein. We installed reactive warheads on chemically prepared ribosylated ubiquitin to generate a set of probes targeting these Legionella enzymes. In vitro tests on recombinant DupA revealed that a vinyl sulfonate warhead was most efficient in covalent complex formation. Mutagenesis and X-ray crystallography approaches were used to identify the site of covalent cross-linking to be an allosteric cysteine residue. The subsequent application of this probe highlights the potential to selectively enrich the Dup enzymes from Legionella-infected cell lysates.

Covalent Probes To Capture Legionella pneumophila Dup Effector Enzymes.,Kloet MS, Mukhopadhyay R, Mukherjee R, Misra M, Jeong M, Talavera Ormeno CMP, Moutsiopoulou A, Tjokrodirijo RTN, van Veelen PA, Shin D, Dikic I, Sapmaz A, Kim RQ, van der Heden van Noort GJ J Am Chem Soc. 2024 Oct 2;146(39):26957-26964. doi: 10.1021/jacs.4c08168. Epub , 2024 Sep 17. PMID:39288007^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.