V5-tagged LIMD1 in HEK293 cells with or without JNK2 activation

The experiment was conducted to identify JNK phosphorylation sites on human LIM domain containing protein-1 (LIMD1). V5-tagged LIMD1 was co-transfected or not with a constitutively active form of JNK2 and purified. The purified proteins were subjected to SDS-PAGE, protease K digestion and MS/MS analysis. The data were searched against Swissprot using Mascot server v2.3. The fixed modification was set to carbamidomethylation of C, and variable modifications were set to oxidation of M, phosphorylation of ST, phosphorylation of Y. Mass values were set as monoisotopic. Peptide mass tolerance was set to +/-10ppm, and MS/MS tolerance was set to +/-0.8Da.