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Identification of NEDD8 sites at endogenous levels of NEDD8 expression upon NEDP1 (SENP8, DEN1) knockout.

ABSTRACT: We developed a strategy that allows the identification of NEDP1 dependent NEDDylation sites under endogenous expression of wild type NEDD8. We combined the use of anti-diGly antibodies that recognise both ubiquitin and NEDD8 modified peptides upon trypsin digestion with short treatment of cells with the ubiquitin E1 inhibitor MLN7243 (UAEi), that dramatically reduces ubiquitin but not NEDD8 modification. By eliminating the majority of ubiquitin-derived diGly peptides upon MLN7243 treatment, we would be able to quantify NEDP1 dependent diGly peptides. Extracts from parental and NEDP1 knockout HCT116 cells both treated with UAEi were used for the isolation of diGly modified peptides and mass spectrometry analysis.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell, Colon

DISEASE(S): Colon Cancer

SUBMITTER: Dimitris Xirodimas

LAB HEAD: Dimitris Xirodimas

PROVIDER: PXD040466 | Pride | 2023-05-10

REPOSITORIES: Pride

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Targeting the NEDP1 enzyme to ameliorate ALS phenotypes through stress granule disassembly.

Kassouf Toufic T Shrivastava Rohit R Meszka Igor I Bailly Aymeric A Polanowska Jolanta J Trauchessec Helene H Mandrioli Jessica J Carra Serena S Xirodimas Dimitris P DP

Science advances 20230331 13

The elimination of aberrant inclusions is regarded as a therapeutic approach in neurodegeneration. In amyotrophic lateral sclerosis (ALS), mutations in proteins found within cytoplasmic condensates called stress granules (SGs) are linked to the formation of pathological SGs, aberrant protein inclusions, and neuronal toxicity. We found that inhibition of NEDP1, the enzyme that processes/deconjugates the ubiquitin-like molecule NEDD8, promotes the disassembly of physiological and pathological SGs. ...[more]

PMID: 37000881

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Proteome-wide identification of NEDD8 modification sites reveals distinct proteomes for canonical and atypical NEDDylation

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2021-02-10 | PXD022812 | Pride

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2009-03-23 | E-GEOD-14088 | biostudies-arrayexpress

E2-RING expansion of the NEDD8 cascade confers specificity to cullin modification

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Hat1 WT and Hat1 KO MEFs analyzed to identify sites of acetylation that are dependent on Hat1

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