TY  - INPR
A1  - Kloet, Max S.
A1  - Mukhopadhyay, Rishov
A1  - Mukherjee, Rukmini
A1  - Misra, Mohit
A1  - Jeong, Minwoo
A1  - Talavera Ormeño, Cami M. P.
A1  - Moutsiopoulou, Angeliki
A1  - Tjokrodirijo, Rayman T. N.
A1  - Veelen, Peter A. van
A1  - Shin, Donghyuk
A1  - Đikić, Ivan
A1  - Sapmaz, Aysegul
A1  - Kim, Robbert Q.
A1  - Heden van Noort, Gerbrand J. van der
T1  - Development of covalent probes to capture Legionella pneumophila effector enzymes
T2  - bioRxiv
N2  - Upon infection of host cells, Legionella pneumophila releases a multitude of effector enzymes into the cells cytoplasm that hijack a plethora of cellular activities, including the hosts ubiquitination pathways. Effectors belonging to the SidE-family are involved in non-canonical 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 hydrolyse 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 crosslinking 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.
Y1  - 2024
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/83415
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-834154
UR  - https://www.biorxiv.org/content/10.1101/2024.03.19.585531v1
IS  - 2024.03.19.585531 Version 1
PB  - bioRxiv
ER  -