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Quantitative ubiquitinome of VCPIP1

ABSTRACT: Quantitative ubiquitinome was performed in NRCMs transfected with empty vector (EV) or VCPIP1WT and challenged with Ang II. Each group was represented by a pooled sample from three biological replicates. The quantitative ubiquitinome was carried out by PTM Bio (Hangzhou, China). Cell samples were lysed in lysis buffer (8 M urea, 1% protease inhibitor cocktail). To enrich modified peptides, tryptic peptides were incubated with anti-diglycine lysine (K-ε-GG) antibody beads (PTM-1104). Peptides were separated at a constant flow rate of 450 nl/min on an Easy-nLC1000 UHPLC system (Bruker Daltonics). The peptides were subjected to capillary source followed by the timsTOF Pro mass spectrometry. The electrospray voltage applied was 1.8 kV. Precursors and fragments were analyzed at the TOF detector, with a MS/MS scan range from 100-1700. The timsTOF Pro was operated in parallel accumulation serial fragmentation (PASEF) mode. Precursors with charge states 0-5 were selected for fragmentation, and 10PASEF-MS/MS scans were acquired per cycle. The dynamic exclusion was set to 24 s. The resulting MS/MS data were processed using MaxQuant search engine (v.1.6.15.0). False discovery rate (FDR) was adjusted to < 1%.14196 ubiquitinated peptides, 14466 ubiquitinated sites, 7641 comparable sites, and 2235 comparable proteins were identified.

ORGANISM(S): Rattus Norvegicus

SUBMITTER: Guang Liang

PROVIDER: PXD076364 | iProX | Tue Mar 31 00:00:00 GMT+01:00 2026

REPOSITORIES: iProX

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