Project description:GFP-tagged viral peptide sequences and their controls were used to pull down interactors from human (uninfected) or green monkey (viral infected) cell lysate. After sample preparation, digestion and bottom-up LC-MS/MS analysis, the enriched proteins were identified.
Project description:Cell lysates from HEK293 cells individually transfected with GFP or GFP-CYYR1 were purified on GFP-Trap affinity resin and analyzed by quantitative label-free mass spectrometry. Experiments were performed in 5 biological replicates for each condition.
Project description:This entry contains proteome data extracted from co-immunoprecipitation of engineered H3F3A locus fused to GFP in HEK293 cell line, followed by PAGE separation, in-gel proteolytic digestion and LC-MS/MS identification.
Project description:In this study, we pull-down human Talin1-GFP and GFP from U2OS cells plated on fibronectin. Binding partners were then analysed using mass-spectrometry.
Project description:To explore the possible neddylation sites, GFP-IRF7 and His-NEDD8 were co-overexpressed in HEK293 cell line and GFP was immunoprecipitated, followed by SDS-PAGE and silver staining. The band detected was subjected to LC–MS/MS.
Project description:This dataset describes a proteomic analysis of proteins co-immunoprecipitated with functional GFP-tagged PILS2, PILS3, and PILS6 fusion proteins in Arabidopsis thaliana roots. PILS (PIN-LIKES) proteins are auxin transport facilitators that localize to the endoplasmic reticulum (ER). To identify proteins that associate with PILS2, PILS3, and PILS6 under physiological conditions, we performed GFP-based immunoprecipitation (IP) followed by mass spectrometry (MS). Protein extracts were prepared from root tissues of Arabidopsis Col-0 (wild-type) and PILS2-, PILS3-, and PILS6-overexpressing transgenic lines expressing N- or C-terminal GFP fusion proteins. Plant material was ground in liquid nitrogen, and total proteins were extracted using aTris Buffer containing 1% CHAPS, 20 mM Tris-HCl (pH 7.5), 10 mM MgCl₂, 15 mM EGTA, 75 mM NaCl, 1 mM DTT, 0.1% Tween-20, and protease inhibitors. Extracts were centrifuged, and the supernatants were quantified using the Bradford assay. 2mg of protein were subjected to immunoprecipitation using anti-GFP magnetic beads, Col-0 (non-tagged) extracts were used as negative controls. Co-immunoprecipitated proteins were then processed and analyzed by mass spectrometry.
Project description:To determine which cellular proteins interacts with the viral MRV protein μ2 and its mutant μ2-P208S, GFP-tagged constructions of μ2 and μ2-P208S in both C- and N- terminus were transfected in 293T cells alongside control GFP alone for 24h, and submitted to GFP pulldown before IP were prepared for MS analysis.