Notes: A novel method for affinity purification mass spectrometry (AP-MS) is described using Magne® HaloTag® Beads. The Sin3 histone deacetylase complex subunits were tagged, and interactions with the remaining components were detected. Cellular lysates were incubated with Magne® HaloTag® Beads to enrich for tagged proteins. Together, an interaction network for the chromatin remodeling complex is described. (5100)

Notes: Pleckstrin-homology N1 (PLEKHN1) is involved in reactive oxygen species (ROS) -induced apoptosis, and knockdown of PLEKHN1 in human colon cancer cells lead to increased cell survival. Interestingly, PLEKHN1 expression was absent in cancer patient samples, possibly explaining decreased apoptosis in cancer cells. Magne® HaloTag® Beads were used to pull down PLEKHN1-interacting factors, and an interaction with Bid, a pro-apoptotic protein, was found. PLEKHN1 knockout cells showed loss of Bid localization at the mitochondrial outer membrane and loss of Bak-Bax oligomerization. (5101)