Visvanathan, Ramya and Utsuki, Tadanobu and Beck, Daniel E. and Clayton, W. Brent and Lendy, Emma and Sun, Kuai-lin and Liu, Yinghui and Hering, Kirk W. and Mesecar, Andrew and Zhang, Zhong-Yin and Putt, Karson S. and Kim, Cheorl-Ho (2024) A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes. PLOS ONE, 19 (3). e0299541. ISSN 1932-6203
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Abstract
The activities of the phospholipase C gamma (PLCγ) 1 and 2 enzymes are essential for numerous cellular processes. Unsurprisingly, dysregulation of PLCγ1 or PLCγ2 activity is associated with multiple maladies including immune disorders, cancers, and neurodegenerative diseases. Therefore, the modulation of either of these two enzymes has been suggested as a therapeutic strategy to combat these diseases. To aid in the discovery of PLCγ family enzyme modulators that could be developed into therapeutic agents, we have synthesized a high-throughput screening-amenable micellular fluorogenic substrate called C16CF3-coumarin. Herein, the ability of PLCγ1 and PLCγ2 to enzymatically process C16CF3-coumarin was confirmed, the micellular assay conditions were optimized, and the kinetics of the reaction were determined. A proof-of-principle pilot screen of the Library of Pharmacologically Active Compounds 1280 (LOPAC1280) was performed. This new substrate allows for an additional screening methodology to identify modulators of the PLCγ family of enzymes.
Item Type: | Article |
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Subjects: | Eprint Open STM Press > Biological Science |
Depositing User: | Unnamed user with email admin@eprint.openstmpress.com |
Date Deposited: | 30 Mar 2024 11:55 |
Last Modified: | 30 Mar 2024 11:55 |
URI: | http://library.go4manusub.com/id/eprint/2089 |