NEDD8 subtrates from NEDP1 knockout cells

ABSTRACT: The aim of the study was to identify proteins that are modified by NEDD8. The NEDD8 specific protease NEDP1/SENP8 was deleted from HEK 293 cells via Crispr/CAS9 gene editing to allow for the accumulation of NEDD8 modified proteins. Lysates from NEDP1 KO cells were then enriched via pulldown with a catalytically inactivated NEDP1 (C162A) fused to the HALO protein. For a negative control a mutated NEDP1 (DAGC) with reduced binding to NEDD8 was also fused to the HALO protein for pulldown. Pulldowns were resolved by SDS-PAGE and bands were excised and subjected to in gel trypsin digestion followed by mass spectrometry analysis.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Matthew Keuss  

LAB HEAD: Thimo Kurz

PROVIDER: PXD011928 | Pride | 2019-02-20

REPOSITORIES: Pride