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Ubiquitin substrates were identified in SILAC-labeled H1299 wild-type and MPND KO cells using LC-MS/MS.

ABSTRACT: Using H1299 wild-type cells and their MPND-knockout derivatives as research subjects, this project employs Stable Isotope Labeling by Amino Acids in Cell Culture (SILAC) for metabolic labeling of the two cell groups. Combined with ubiquitinated peptide-specific enrichment and nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS) techniques, we aim to establish a quantitative expression profile of ubiquitin proteins in these cells. Subsequent data retrieval, quantitative analysis, and differential screening will be performed using software such as MaxQuant to clarify the differential expression characteristics of ubiquitin proteins in H1299 cells following MPND knockout, including up-regulated and down-regulated ubiquitin proteins as well as specific ubiquitination modification sites. Furthermore, bioinformatics analyses—including Gene Ontology (GO) functional enrichment, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and protein-protein interaction (PPI) network construction—will be utilized to predict the signaling pathways involved in these differential ubiquitin proteins and the potential downstream substrates regulated by MPND.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Dawei Li

LAB HEAD: Dawei Li

PROVIDER: PXD074990 | Pride | 2026-05-15

REPOSITORIES: Pride

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