A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes.
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 modulati...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2024-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0299541&type=printable |
| _version_ | 1797224038559383552 |
|---|---|
| author | Ramya Visvanathan Tadanobu Utsuki Daniel E Beck W Brent Clayton Emma Lendy Kuai-Lin Sun Yinghui Liu Kirk W Hering Andrew Mesecar Zhong-Yin Zhang Karson S Putt |
| author_facet | Ramya Visvanathan Tadanobu Utsuki Daniel E Beck W Brent Clayton Emma Lendy Kuai-Lin Sun Yinghui Liu Kirk W Hering Andrew Mesecar Zhong-Yin Zhang Karson S Putt |
| author_sort | Ramya Visvanathan |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-24T13:46:46Z |
| format | Article |
| id | doaj.art-07330adcb09c40d7bd2c75877bb72103 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-24T13:46:46Z |
| publishDate | 2024-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-07330adcb09c40d7bd2c75877bb721032024-04-04T05:34:30ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-01193e029954110.1371/journal.pone.0299541A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes.Ramya VisvanathanTadanobu UtsukiDaniel E BeckW Brent ClaytonEmma LendyKuai-Lin SunYinghui LiuKirk W HeringAndrew MesecarZhong-Yin ZhangKarson S PuttThe 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.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0299541&type=printable |
| spellingShingle | Ramya Visvanathan Tadanobu Utsuki Daniel E Beck W Brent Clayton Emma Lendy Kuai-Lin Sun Yinghui Liu Kirk W Hering Andrew Mesecar Zhong-Yin Zhang Karson S Putt A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes. PLoS ONE |
| title | A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes. |
| title_full | A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes. |
| title_fullStr | A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes. |
| title_full_unstemmed | A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes. |
| title_short | A novel micellular fluorogenic substrate for quantitating the activity of 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma (PLCγ) enzymes. |
| title_sort | novel micellular fluorogenic substrate for quantitating the activity of 1 phosphatidylinositol 4 5 bisphosphate phosphodiesterase gamma plcγ enzymes |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0299541&type=printable |
| work_keys_str_mv | AT ramyavisvanathan anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT tadanobuutsuki anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT danielebeck anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT wbrentclayton anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT emmalendy anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT kuailinsun anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT yinghuiliu anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT kirkwhering anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT andrewmesecar anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT zhongyinzhang anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT karsonsputt anovelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT ramyavisvanathan novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT tadanobuutsuki novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT danielebeck novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT wbrentclayton novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT emmalendy novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT kuailinsun novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT yinghuiliu novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT kirkwhering novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT andrewmesecar novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT zhongyinzhang novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes AT karsonsputt novelmicellularfluorogenicsubstrateforquantitatingtheactivityof1phosphatidylinositol45bisphosphatephosphodiesterasegammaplcgenzymes |