Identification of the NEDDylated proteome by Nedd8-specific K-GG mass spectrometry uncovers modulation of Actin dynamics by neddylated Cofilin
Ontology highlight
ABSTRACT: Neddylation is the post-translational protein modification that is most closely related to
ubiquitination. The Ubiquitin-like protein NEDD8 is mostly studied for its role in activating
the Cullin-RING E3 Ubiquitin ligases, however little is known about other NEDD8 targets.
We developed serial NEDD8-Ubiquitin Substrate Profiling (sNUSP), a method that
employs Nedd8-R74K knock-in cells allowing discrimination of endogenous NEDD8- and
Ubiquitin-modification sites by mass spectrometry after Lys-C digestion and K-GG-
peptide enrichment. Using sNUSP, we identified 607 neddylation sites dynamically
regulated by the neddylation inhibitor MLN4924 and the de-neddylating enzyme
NEDP1/SENP8. Among the candidates, we characterized lysine 112 (K112) of the Actin
regulator Cofilin as a novel neddylation event. Global inhibition of neddylation in
developing neurons leads to cytoskeletal defects, altered Actin dynamics and neurite
growth impairments and site-specific neddylation of Cofilin at K112 regulates neurite
outgrowth. These data show that Cofilin neddylation contributes to the regulation of
neuronal Actin dynamics.
INSTRUMENT(S): LTQ Orbitrap
ORGANISM(S): Homo Sapiens (ncbitaxon:9606)
SUBMITTER: erik verschueren Donald Kirkpatrick
PROVIDER: MSV000084683 | MassIVE | Thu Dec 12 20:59:00 GMT 2019
REPOSITORIES: MassIVE
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