Human Sumoylome (SUMO-2) upon bacterial infection

Given the role of SUMOylation in pathogenic infection, we wanted to evaluate the changes to the host SUMO-2 subproteome during Shigella infection. HeLa cells overexpressing TAP-SUMO-2 or TAP-empty were used in conjunction with SILAC (Stable Isotope Labeling of Amino acids in Culture) (Golebiowski et al, 2009, 2010). Cells were grown in Dulbecco’s modified Eagle’s medium except that L-arginine and L-lysine were replaced with stable isotope (SILAC) forms depending on the treatment. Medium was supplemented with 10% dialyzed fetal calf serum. SILAC experiment compared TAP-containing cells (Lys0 and Arg0) with invasive Shigella flexneri (M90T) infected TAP-SUMO-2-containing cells (4,4,5,5-D4-lysine, Lys4, and 13C6- arginine, Arg6) and TAP-SUMO-2-containing cells infected with (mxiD) non-invasive strain of Shigella (13C6 15N2-lysine, Lys8, and 13C6 15N4 -arginine, Arg10).