Project description:Mass spectrometry study of interactor binding to the EVH1 domain of VASP in mature and immature dendritic cells. We performed mass spectrometry study using a specially designed inhibitor that recognizes this characterized motif with high affinity and analyzed the digested peptides by LC-MS. For both, iDCs and mDCs, two independent experiments were carried out (N=2). As control for the specificity of the identified interactions an additional two independent experiments (N=2) were carried out for both, iDCs and mDCs, where a specific inhibitor of the EVH1 domain was added to outcompete bona fide EVH1 domain interactions.
Project description:LC-MS/MS analysis of Control, PAR1-stimulated and PAR4-stimulated platelet lysate digests. Control lysate digest peptides were labeled with "light", PAR1 lysate digest peptides were labeled with "intermediate" and PAR4 lysate digest peptides were labeled with "high" dimethyl labeled. Labeled digests were mixed in a 1:1:1 ratio, separated by SCX and the resulting (phosphopeptide-enriched) peptide fractions were analyzed by LC-MS/MS.