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Membrane proteome analysis of sorted splenic CD4+ T cells from SRBCs-immunized Rabggta-icKO, Ggps1-icKO and WT mice

ABSTRACT: In order to explore the effects of Rabggta or Ggps1 on the membrane proteome of T cells, sorted CD4+ T cells of SRBCs-immunized Rabggta-icKO, Ggps1-icKO and their WT mice were subjected to membrane protein extraction with standard protocol. Take 50 μg protein and diluted to 100 μL water. Then add 5 μL (0.25 μg/μL) trpsin enzyme and incubate in a 37℃ constant temperature incubator for 16 hours for protein digestion. Then samples were subjected to Nano LC-MS/MS analysis. The raw MS files were analyzed and searched against target protein database based on the species of the samples using MaxQuant. The parameters were set as follows: the protein modifications were carbamidomethylation (C) (fixed), oxidation (M) (variable), Acetyl (N-term) (variable); the enzyme specificity was set to trypsin; the maximum missed cleavages were set to 2; the precursor ion mass tolerance was set to 20 ppm, and MS/MS tolerance was 20 ppm. Only high confident identified peptides were chosen for downstream protein identification analysis. Gene ontology (GO) terms were used to obtain and classify the function information of the DEPs.

ORGANISM(S): Mus Musculus

SUBMITTER: Lai Wang

PROVIDER: PXD062705 | iProX | Mon Apr 07 00:00:00 BST 2025

REPOSITORIES: iProX

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