A membrane-associated, fluorogenic reporter for mammalian phospholipase C isozymes

14Citations
Citations of this article
33Readers
Mendeley users who have this article in their library.

This article is free to access.

Abstract

A diverse group of cell-surface receptors, including many G protein-coupled receptors and receptor tyrosine kinases, activate phospholipase C (PLC) isozymes to hydrolyze phosphatidylinositol 4,5-bisphosphate into the second messengers diacylglycerol and 1,4,5-inositol trisphosphate. Consequently, PLCs control various cellular processes, and their aberrant regulation contributes to many diseases, including cancer, atherosclerosis, and rheumatoid arthritis. Despite the widespread importance of PLCs in human biology and disease, it has been impossible to directly monitor the real-time activation of these enzymes at membranes. To overcome this limitation, here we describe XY-69, a fluorogenic reporter that preferentially partitions into membranes and provides a selective tool for measuring the real-time activity of PLCs as either purified enzymes or in cellular lysates. Indeed, XY-69 faithfully reported the membrane-dependent activation of PLC-3 by Gq. Therefore, XY-69 can replace radioactive phosphatidylinositol 4,5-bisphosphate used in conventional PLC assays and will enable high-throughput screens to identify both orthosteric and allosteric PLC inhibitors. In the future, cell-permeable variants of XY-69 represent promising candidates for reporting the activation of PLCs in live cells with high spatiotemporal resolution.

Cite

CITATION STYLE

APA

Huang, W., Wang, X., Endo-Streeter, S., Barrett, M., Waybright, J., Wohlfeld, C., … Zhang, Q. (2018). A membrane-associated, fluorogenic reporter for mammalian phospholipase C isozymes. Journal of Biological Chemistry, 293(5), 1728–1735. https://doi.org/10.1074/jbc.RA117.000926

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Save time finding and organizing research with Mendeley

Sign up for free