Dataset Information

Identification of Bona Fide Targets of AMPylating Fic-enzymes by Substrate Mediated Covalent Capture

ABSTRACT: Many pathogenic bacteria utilize posttranslational modifications to manipulate central components of the host cell functions. Often, the released enzymes belong to the large Fic-family, causing target modification with nucleotide monophosphates. Due to the lack of a generic method for the identification of the Fic-family’s cellular targets, we have established a new approach by combining synthetic thiol-reactive nucleotide derivatives (TReNDs) with recombinantly produced Fic-enzymes containing strategically placed cysteines in their active sites, thereby producing reactive binary FicTReND probes for covalent substrate capture. The binary probes successfully capture their targets from complex lysates and permit their identification by mass spectrometry.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Christoph Krisp

LAB HEAD: Azmelt tyen

PROVIDER: PXD015599 | Pride | 2020-07-09

REPOSITORIES: Pride

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Identification of targets of AMPylating Fic enzymes by co-substrate-mediated covalent capture.

Gulen Burak B Rosselin Marie M Fauser Joel J Albers Michael F MF Pett Christian C Krisp Christoph C Pogenberg Vivian V Schlüter Hartmut H Hedberg Christian C Itzen Aymelt A

Nature chemistry 20200706 8

Various pathogenic bacteria use post-translational modifications to manipulate the central components of host cell functions. Many of the enzymes released by these bacteria belong to the large Fic family, which modify targets with nucleotide monophosphates. The lack of a generic method for identifying the cellular targets of Fic family enzymes hinders investigation of their role and the effect of the post-translational modification. Here, we establish an approach that uses reactive co-substrate- ...[more]

PMID: 32632184

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