Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptor
Background: Gintonin is a ginseng-derived exogenous G-protein-coupled lysophosphatidic acid (LPA) receptor ligand. Gintonin exerts its neuronal and non-neuronal in vitro and in vivo effects through LPA receptor subtypes. However, it is unknown whether gintonin can bind to the plasma membrane of cell...
| Main Authors: | , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2022-05-01
|
| Series: | Journal of Ginseng Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1226845321001573 |
| _version_ | 1818242438472400896 |
|---|---|
| author | Sun-Hye Choi Ra Mi Lee Han-Sung Cho Sung Hee Hwang Hong-Ik Hwang Hyewhon Rhim Hyoung-Chun Kim Do-Geun Kim Ik-Hyun Cho Seung-Yeol Nah |
| author_facet | Sun-Hye Choi Ra Mi Lee Han-Sung Cho Sung Hee Hwang Hong-Ik Hwang Hyewhon Rhim Hyoung-Chun Kim Do-Geun Kim Ik-Hyun Cho Seung-Yeol Nah |
| author_sort | Sun-Hye Choi |
| collection | DOAJ |
| description | Background: Gintonin is a ginseng-derived exogenous G-protein-coupled lysophosphatidic acid (LPA) receptor ligand. Gintonin exerts its neuronal and non-neuronal in vitro and in vivo effects through LPA receptor subtypes. However, it is unknown whether gintonin can bind to the plasma membrane of cells and can transactivate the epidermal growth factor (EGF) receptor. In the present study, we examined whether gintonin-biotin conjugates directly bound to LPA receptors and transactivated the EGF receptor. Methods: We designed gintonin-biotin conjugates through gintonin biotinylation and examined whether gintonin-biotin conjugate binding sites co-localized with the LPA receptor subtype binding sites. We further examined whether gintonin-biotin transactivated the EGF receptor via LPA receptor regulation via phosphor-EGF and cell migration assays. Results: Gintonin-biotin conjugates elicit [Ca2+]i transient similar to that observed with unbiotinylated gintonin in cultured PC3 cells, suggesting that biotinylation does not affect physiological activity of gintonin. We proved that gintonin-biotin conjugate binding sites co-localized with the LPA1/6 receptor binding sites. Gintonin-biotin binding to the LPA1 receptor transactivates the epidermal growth factor (EGF) receptor through phosphorylation, while the LPA1/3 receptor antagonist, Ki16425, blocked phosphorylation of the EGF receptor. Additionally, an EGF receptor inhibitor AG1478 blocked gintonin-biotin conjugate-mediated cell migration. Conclusions: We observed the binding between ginseng-derived gintonin and the plasma membrane target proteins corresponding to the LPA1/6 receptor subtypes. Moreover, gintonin transactivated EGF receptors via LPA receptor regulation. Our results suggest that gintonin directly binds to the LPA receptor subtypes and transactivates the EGF receptor. It may explain the molecular basis of ginseng physiology/pharmacology in biological systems. |
| first_indexed | 2024-12-12T13:45:14Z |
| format | Article |
| id | doaj.art-f94a51922bf14620a9cecae6836e7e55 |
| institution | Directory Open Access Journal |
| issn | 1226-8453 |
| language | English |
| last_indexed | 2024-12-12T13:45:14Z |
| publishDate | 2022-05-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Ginseng Research |
| spelling | doaj.art-f94a51922bf14620a9cecae6836e7e552022-12-22T00:22:42ZengElsevierJournal of Ginseng Research1226-84532022-05-01463348356Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptorSun-Hye Choi0Ra Mi Lee1Han-Sung Cho2Sung Hee Hwang3Hong-Ik Hwang4Hyewhon Rhim5Hyoung-Chun Kim6Do-Geun Kim7Ik-Hyun Cho8Seung-Yeol Nah9Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University, Seoul, Republic of KoreaGinsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University, Seoul, Republic of KoreaGinsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University, Seoul, Republic of KoreaDepartment of Pharmaceutical Engineering, College of Health Sciences, Sangji University, Wonju, Republic of KoreaCenter for Neuroscience, Korea Institute of Science and Technology, Seoul, Republic of KoreaCenter for Neuroscience, Korea Institute of Science and Technology, Seoul, Republic of KoreaNeuropsychopharmacology and Toxicology Program, College of Pharmacy, Kangwon National University, Chunchon, Republic of KoreaDementia Research Group, Korea Brain Research Institute, Daegu, Republic of KoreaDepartment of Science in Korean Medicine, Brain Korea 21 Plus Program, Department of Conversions Medical Science, Graduate School, Kyung Hee University, Seoul, Republic of KoreaGinsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University, Seoul, Republic of Korea; Corresponding author. Ginsentology Research Laboratory and Department of Physiology, College of Veterinary Medicine, Konkuk University, Seoul, 05029, Republic of Korea.Background: Gintonin is a ginseng-derived exogenous G-protein-coupled lysophosphatidic acid (LPA) receptor ligand. Gintonin exerts its neuronal and non-neuronal in vitro and in vivo effects through LPA receptor subtypes. However, it is unknown whether gintonin can bind to the plasma membrane of cells and can transactivate the epidermal growth factor (EGF) receptor. In the present study, we examined whether gintonin-biotin conjugates directly bound to LPA receptors and transactivated the EGF receptor. Methods: We designed gintonin-biotin conjugates through gintonin biotinylation and examined whether gintonin-biotin conjugate binding sites co-localized with the LPA receptor subtype binding sites. We further examined whether gintonin-biotin transactivated the EGF receptor via LPA receptor regulation via phosphor-EGF and cell migration assays. Results: Gintonin-biotin conjugates elicit [Ca2+]i transient similar to that observed with unbiotinylated gintonin in cultured PC3 cells, suggesting that biotinylation does not affect physiological activity of gintonin. We proved that gintonin-biotin conjugate binding sites co-localized with the LPA1/6 receptor binding sites. Gintonin-biotin binding to the LPA1 receptor transactivates the epidermal growth factor (EGF) receptor through phosphorylation, while the LPA1/3 receptor antagonist, Ki16425, blocked phosphorylation of the EGF receptor. Additionally, an EGF receptor inhibitor AG1478 blocked gintonin-biotin conjugate-mediated cell migration. Conclusions: We observed the binding between ginseng-derived gintonin and the plasma membrane target proteins corresponding to the LPA1/6 receptor subtypes. Moreover, gintonin transactivated EGF receptors via LPA receptor regulation. Our results suggest that gintonin directly binds to the LPA receptor subtypes and transactivates the EGF receptor. It may explain the molecular basis of ginseng physiology/pharmacology in biological systems.http://www.sciencedirect.com/science/article/pii/S1226845321001573GintoninGintonin-biotin conjugatesLPA receptor co-localizationEGF receptorTransactivation |
| spellingShingle | Sun-Hye Choi Ra Mi Lee Han-Sung Cho Sung Hee Hwang Hong-Ik Hwang Hyewhon Rhim Hyoung-Chun Kim Do-Geun Kim Ik-Hyun Cho Seung-Yeol Nah Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptor Journal of Ginseng Research Gintonin Gintonin-biotin conjugates LPA receptor co-localization EGF receptor Transactivation |
| title | Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptor |
| title_full | Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptor |
| title_fullStr | Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptor |
| title_full_unstemmed | Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptor |
| title_short | Visualization of the binding between gintonin, a Panax ginseng-derived LPA receptor ligand, and the LPA receptor subtypes and transactivation of the EGF receptor |
| title_sort | visualization of the binding between gintonin a panax ginseng derived lpa receptor ligand and the lpa receptor subtypes and transactivation of the egf receptor |
| topic | Gintonin Gintonin-biotin conjugates LPA receptor co-localization EGF receptor Transactivation |
| url | http://www.sciencedirect.com/science/article/pii/S1226845321001573 |
| work_keys_str_mv | AT sunhyechoi visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT ramilee visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT hansungcho visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT sungheehwang visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT hongikhwang visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT hyewhonrhim visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT hyoungchunkim visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT dogeunkim visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT ikhyuncho visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor AT seungyeolnah visualizationofthebindingbetweengintoninapanaxginsengderivedlpareceptorligandandthelpareceptorsubtypesandtransactivationoftheegfreceptor |