Identification of Novel Death-Associated Protein Kinase 2 Interaction Partners by Proteomic Screening Coupled with Bimolecular Fluorescence Complementation.

摘要: Death-associated protein kinase 2 (DAPK2) is a Ca(2+)/calmodulin-dependent Ser/Thr that possesses tumor-suppressive functions and regulates programmed cell death, autophagy, oxidative stress, hematopoiesis, motility. As only few binding partners of DAPK2 have been determined, the molecular mechanisms governing these biological are largely unknown. We report identification 180 potential interaction by affinity purification-coupled mass spectrometry, 12 which known DAPK proteins. A small subset established proteins detected in this screen was further investigated bimolecular fluorescence complementation (BiFC) assays, method to visualize interactions living cells. These experiments revealed α-actinin-1 14-3-3-β novel partners. The with localized at plasma membrane, resulting massive membrane blebbing reduced cellular motility, whereas cytoplasm, no impact on blebbing, or viability. Our results therefore suggest effector influenced protein's subcellular localization highlight utility combining spectrometry screening identify characterize protein-protein interactions.