title: Monitoring Inositol-Specific Phospholipase C Activity Using a Phospholipid FlashPlate(R).
abstract: Inositol-specific phospholipase Cs(PLCs) are a group of enzymes involved in the signal transduction pathway of many plasma membrane receptor mediated events. We developed a modified solid surface to capture [(3)H] PIP(2) onto the Basic FlashPlate(R) in order to monitor PLC activity. Our results clearly demonstrate the utility of [(3)H] PIP(2)-Coated Phospholipid FlashPlate(R) microtiter plates for assessing PLC activity for HTS of receptor-coupled functional assays. The results show that PLC activity can be measured easily from a variety of sources including purified recombinant enzyme preparations, crude HL60 cell lysates and permeabilized A431 human carcinoma cells. Moreover, this format provides a surface comparable to that used for classical solution based radiolabeled mixed phospholipid micelle studies and illustrates the feasibility of this assay for measuring PLC activation in a variety of different drug screening assays.
authors.affiliation: NEN Life Science Products, Boston, MA.
authors.initials: TR
authors.lastname: Mullinax
authors.initials: G
authors.lastname: Henrich
authors.initials: P
authors.lastname: Kasila
authors.initials: DG
authors.lastname: Ahern
authors.initials: EA
authors.lastname: Wenske
authors.initials: C
authors.lastname: Hou
authors.initials: D
authors.lastname: Argentieri
authors.initials: ME
authors.lastname: Bembenek