Screening and verification for proteins that interact with leucine aminopeptidase of Taenia pisiformis using a yeast two-hybrid system.

摘要: Leucine aminopeptidase of Taenia pisiformis (TpLAP) belonging to the M17 peptidase family has been implicated as a stage-differentially expressed protein in adult stage T. pisiformis. In order further dissect biological functions TpLAP growth and development worms, TpLAP-interacting partners were investigated. this study, yeast two-hybrid (Y2H) cDNA library from was constructed. Using pGBKT7-TpLAP bait, proteins interacting with screened by Y2H system positive preys sequenced analyzed using Basic Local Alignment Search Tool (BLAST). Our results showed that six genuine proteins, including LAP, dynein light chain (DLC), SUMO-conjugating enzyme (UBC9), histone-lysine n-methyltransferase, trans-acting transcriptional, one unknown protein, identified via assay. Furthermore, interaction between UBC9 (TpUBC9), an important involved SUMOylation pathway, validated one-to-one assay, co-immunoprecipitation, confocal analysis. These findings provide deeper understanding offer first clue may act novel SUMOylated substrate, suggesting SUMO modification pathway plays role regulation worm development.