Proteome-wide identification of ISG15 sites targeted by SARS-CoV-2 PLpro

ABSTRACT: Here, we applied an ISG15-GlyGly peptidomics approach on HeLa cells producing high levels of ISGylated proteins upon stimulation with IFN-α. After obtaining cellular lysates from wild-type and Isg15-/-cells, the samples were incubated with recombinant wild-type or mutant PLpro to catalyze deISGylation of host cellular proteins. After incubation with PLpro, we continued with trypsin digestion to generate diglycine-modified peptides that were subsequently enriched by immunoprecipitation and identified and quantified by LC-MS/MS. By comparing sites across all four conditions, we uncovered 118 ISGylation sites on 95 proteins as targets of the SARS-CoV-2 protease PLpro.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Hela Cell

SUBMITTER: Denzel Eggermont  

LAB HEAD: Francis Impens

PROVIDER: PXD026748 | Pride | 2021-10-12

REPOSITORIES: Pride