Arabidopsis SKD1 Proteom LC-MSMS

We investigated the heat-dependent in-vivo interactome of Arabidopsis thaliana SKD1 using a transgenic 35S::GFP-SKD1 line. Potential interactors of GFP-SKD1 were co-immunoprecipitated from cell extracts of untreated or heat-treated rosette leaves and analyzed by mass spectrometry. A line overexpressing free YFP (35S::YFP) was used as a negative control. For each genotype and condition, proteins of three biological replicates were analyzed. An in-solution digest was performed on the beads, and peptides were subjected to liquid chromatography and tandem mass spectrometry (LC-MS/MS, Dr. S. Müller, CECAD/CMMC Proteomics Facility Cologne).