Chemical reporters for exploring ADP-ribosylation and AMPylation at the host–pathogen interface

Abstract

Bacterial pathogens secrete protein toxins and effectors that hijack metabolites to covalently modify key host proteins and interfere with their function during infection. Adenosine metabolites, such as nicotinamide adenine dinucleotide (NAD) and adenosine triphosphate (ATP), have in particular been co-opted by these secreted virulence factors to reprogram host pathways. While some host targets for secreted virulence factors have been identified, other toxin and effector substrates have been elusive, which require new methods for their characterization. In this review, we focus on chemical reporters based on NAD and ATP that should facilitate the discovery and characterization of adenosine diphosphate (ADP)-ribosylation and adenylylation/AMPylation in bacterial pathogenesis and cell biology.

Chemical proteomics reveals ADP-ribosylation of small GTPases during oxidative stress

“…ADP-ribosylation also plays a prominent role in host-pathogen interactions 11 . Notably, diphtheria toxin catalyzed ADP-ribosylation, which was discovered shortly after histone ADP-ribosylation, inhibits the function of mammalian elongation factor-2 (EF-2) 12 .…”

“…Alternatively, the bacterial effector ExoS from Pseudomonas aeruginosa can ADP-ribosylated Ras and inhibit its GTPase activity 13 . Other secreted bacterial toxins or effectors also encode ADP-ribosyltransferase activity, but their host protein targets have not been fully characterized 11,14 . Despite the association with human diseases and infections, the precise functions of ADP-ribosylation in many cellular processes are still unknown due to challenges in the biochemical analysis of this complex and dynamic posttranslational modification.…”

High-throughput identification of proteins with AMPylation using self-assembled human protein (NAPPA) microarrays

“…The assay using bioorthogonal chemistry is also non-radioactive, robust and sensitive. The specificity and efficiency of this method, which are far superior to typical antibody-mediated interactions, and the smaller steric profiles of the alkyne and azide groups compared with common protein tags, made this the first choice for the development of a high-throughput AMPylation screening platform on NAPPA arrays 10,25 .…”

rAMPing Up Stress Signaling: Protein AMPylation in Metazoans

“…We shall not cover eukaryotic adenylyl transferases that modify small, non-peptide molecules i.e. nicotinamide mononucleotide adenylyltransferase (reviewed in (Petrelli et al, 2011)), bacterial AMPylases that modify endogenous bacterial targets (reviewed in (Garcia-Pino et al, 2014, Harms et al, 2016, Itzen et al, 2011, Woolery et al, 2010)) or methods to study protein AMPylation (reviewed in (Hedberg and Itzen, 2015, Müller et al, 2014, Westcott and Hang, 2014). …”