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To resolve the controversy about messengers regulating KCNQion channels during phospholipase Cmediated suppressionof current, we designed translocatable enzymes that quicklyalter the phosphoinositide composition of the plasma membraneafter application of a chemical cue. The KCNQ current fallsrapidly to zero when phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2or PI(4,5)P2] is depleted without changing Ca2+, diacylglycerol,or inositol 1,4,5-trisphosphate. Current rises by 30% when PI(4,5)P2is overproduced and does not change when phosphatidylinositol3,4,5-trisphosphate is raised. Hence, the depletion of PI(4,5)P2suffices to suppress current fully, and other second messengersare not needed. Our approach is ideally suited to study biologicalsignaling networks involving membrane phosphoinositides.
1 Department of Physiology and Biophysics, University of Washington School of Medicine, Seattle, WA 98195, USA. 2 Department of Molecular Pharmacology, Stanford University, Clark Center, 318 Campus Drive, Stanford, CA 94305, USA.
* These authors contributed equally to this work.
To whom correspondence should be addressed. E-mail: hille{at}u.washington.edu
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