FIGURE 6.

Identification of TbNedd8-conjugated and -associated proteins by affinity purification and mass spectrometry. A, procyclic cells expressing PTP-tagged TbNedd8 were lysed by sonication, and the cell lysate was used for tandem affinity purification. The final EGTA/EDTA eluate was loaded onto SDS-PAGE and stained with Coomassie Blue. Arrows indicate the protein bands that were analyzed by LC-MS/MS. B, immunoprecipitation of 3HA-TbNedd8-conjugated and -associated proteins for LC-MS/MS. Control cells and cells expressing 3HA-TbNedd8 were lysed and then incubated with anti-HA-agarose beads. C, putative TbNedd8-conjugated and -associated proteins identified by affinity purification were grouped according to their molecular functions. The numbers in the parenthesis indicate the numbers of proteins in each function group.