Our massive effort to characterize protein ADP-ribosylation, phosphorylation, and complexes in macrophages is out in JPR! I’m going to walk you through it. (1/6) https://pubs.acs.org/doi/pdf/10.1021/acs.jproteome.0c00261

MAPCD – Macrophage ADP-ribosylation, Phosphorylation, and Complex Dynamics – is a resource available online and in Supp Table 10. 6,729 proteins; ADP-ribosylation characterized on 2,905 proteins; phosphorylation characterized on 2,669 proteins. (2/6)
https://mapcd.niaid.NIH.gov 

SEC-MS was used to separate proteoforms by size; protein complexes were then identified by WGCNA based on their correlated movement into or out of SEC fractions following stimulation, presenting an analysis method for SEC-MS that does not rely on established databases. (3/6)

Two modules of interest are studied further – one representing the ASK signalosome forming in response to LPS stimulation of macrophages, the other representing a PARP9 centric protein complex that falls apart in response to LPS stimulation. (4/6)

PARP inhibitor treatment of macrophages altered the global interactome dramatically. Considering the ‘scaffolding’ role for poly(ADP-ribose) shown by the @LeungLab and others, this is not surprising but still exciting to see! (5/6)

PARPi treatment also blocked the LPS response in macrophages, at the PTM level (MAP kinase cascade), protein level (TNF release from primary human MΦs), and animal level (endotoxic shock mouse model). ADP-ribosylation is important in the innate immune response! (6/6)