Human Tyrosinase Displayed on the Surface of Chinese Hamster Ovary Cells for Ligand Fishing of Tyrosinase Inhibitors from Medicinal Plants
Ligand fishing is a promising strategy for the screening of active ingredients from complex natural products. In this work, human tyrosinase (hTYR) was displayed on the surface of Chinese hamster ovary (CHO) cells for the first time; it was then used as bait to develop a new method for ligand fishin...
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2024-12-01
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| author | Xiao-Rui Zhai Ming-Jie Li Xiang Yin Ayzohra Ablat Yuan Wang Peng Shu Xun Liao |
| author_facet | Xiao-Rui Zhai Ming-Jie Li Xiang Yin Ayzohra Ablat Yuan Wang Peng Shu Xun Liao |
| author_sort | Xiao-Rui Zhai |
| collection | DOAJ |
| description | Ligand fishing is a promising strategy for the screening of active ingredients from complex natural products. In this work, human tyrosinase (hTYR) was displayed on the surface of Chinese hamster ovary (CHO) cells for the first time; it was then used as bait to develop a new method for ligand fishing. The localization of hTYR on the CHO cell surface was verified by an enzyme activity test and fluorescence microscopy. The displayed tyrosinase (CHO@hTYR) maintained relatively stable enzymatic activity (82.59 ± 2.70%) within 7 days. Furthermore, it can be reused for fishing five times. Guided by the proposed ligand fishing method, four tyrosinase inhibitors, including 4-methoxy-5-methyl coumarin (<b>1</b>), cupressuflavone (<b>2</b>), amentoflavone (<b>3</b>), and 3,4-dimethoxy-5-methyl coumarin (<b>4</b>), were isolated from <i>Alhagi sparsifolia</i>, and the active fraction with low polarity was isolated from <i>Coffea arabica</i>; these two medicinal plants possess skin-lightening potential. All the isolated tyrosinase inhibitors significantly reduced the intracellular tyrosinase activity and melanin level in B16 cells enhanced by α-MSH. Meanwhile, the active fraction (100 μg/mL) from <i>C. arabica</i> exhibited stronger inhibitory effects than the positive controls (α-arbutin and kojic acid) by recovering them to the normal levels. This work demonstrated the promising application of the cell surface display in the field of ligand fishing and is helpful in unveiling the chemical basis of the skin-lightening effect of <i>A. sparsifolia</i> and <i>C. arabica</i>. |
| format | Article |
| id | doaj-art-6c43db201d934b7b9dcd6315912a1894 |
| institution | Kabale University |
| issn | 1420-3049 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj-art-6c43db201d934b7b9dcd6315912a18942025-01-10T13:18:39ZengMDPI AGMolecules1420-30492024-12-013013010.3390/molecules30010030Human Tyrosinase Displayed on the Surface of Chinese Hamster Ovary Cells for Ligand Fishing of Tyrosinase Inhibitors from Medicinal PlantsXiao-Rui Zhai0Ming-Jie Li1Xiang Yin2Ayzohra Ablat3Yuan Wang4Peng Shu5Xun Liao6Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, ChinaHBN Research Institute and Biological Laboratory, Shenzhen Hujia Technology Co., Ltd., Shenzhen 518000, ChinaChengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, ChinaChengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, ChinaHBN Research Institute and Biological Laboratory, Shenzhen Hujia Technology Co., Ltd., Shenzhen 518000, ChinaHBN Research Institute and Biological Laboratory, Shenzhen Hujia Technology Co., Ltd., Shenzhen 518000, ChinaChengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, ChinaLigand fishing is a promising strategy for the screening of active ingredients from complex natural products. In this work, human tyrosinase (hTYR) was displayed on the surface of Chinese hamster ovary (CHO) cells for the first time; it was then used as bait to develop a new method for ligand fishing. The localization of hTYR on the CHO cell surface was verified by an enzyme activity test and fluorescence microscopy. The displayed tyrosinase (CHO@hTYR) maintained relatively stable enzymatic activity (82.59 ± 2.70%) within 7 days. Furthermore, it can be reused for fishing five times. Guided by the proposed ligand fishing method, four tyrosinase inhibitors, including 4-methoxy-5-methyl coumarin (<b>1</b>), cupressuflavone (<b>2</b>), amentoflavone (<b>3</b>), and 3,4-dimethoxy-5-methyl coumarin (<b>4</b>), were isolated from <i>Alhagi sparsifolia</i>, and the active fraction with low polarity was isolated from <i>Coffea arabica</i>; these two medicinal plants possess skin-lightening potential. All the isolated tyrosinase inhibitors significantly reduced the intracellular tyrosinase activity and melanin level in B16 cells enhanced by α-MSH. Meanwhile, the active fraction (100 μg/mL) from <i>C. arabica</i> exhibited stronger inhibitory effects than the positive controls (α-arbutin and kojic acid) by recovering them to the normal levels. This work demonstrated the promising application of the cell surface display in the field of ligand fishing and is helpful in unveiling the chemical basis of the skin-lightening effect of <i>A. sparsifolia</i> and <i>C. arabica</i>.https://www.mdpi.com/1420-3049/30/1/30cell surface displayChinese hamster ovary cellstyrosinase inhibitors<i>Alhagi sparsifolia</i><i>Coffea arabica</i>B16 melanoma cells |
| spellingShingle | Xiao-Rui Zhai Ming-Jie Li Xiang Yin Ayzohra Ablat Yuan Wang Peng Shu Xun Liao Human Tyrosinase Displayed on the Surface of Chinese Hamster Ovary Cells for Ligand Fishing of Tyrosinase Inhibitors from Medicinal Plants Molecules cell surface display Chinese hamster ovary cells tyrosinase inhibitors <i>Alhagi sparsifolia</i> <i>Coffea arabica</i> B16 melanoma cells |
| title | Human Tyrosinase Displayed on the Surface of Chinese Hamster Ovary Cells for Ligand Fishing of Tyrosinase Inhibitors from Medicinal Plants |
| title_full | Human Tyrosinase Displayed on the Surface of Chinese Hamster Ovary Cells for Ligand Fishing of Tyrosinase Inhibitors from Medicinal Plants |
| title_fullStr | Human Tyrosinase Displayed on the Surface of Chinese Hamster Ovary Cells for Ligand Fishing of Tyrosinase Inhibitors from Medicinal Plants |
| title_full_unstemmed | Human Tyrosinase Displayed on the Surface of Chinese Hamster Ovary Cells for Ligand Fishing of Tyrosinase Inhibitors from Medicinal Plants |
| title_short | Human Tyrosinase Displayed on the Surface of Chinese Hamster Ovary Cells for Ligand Fishing of Tyrosinase Inhibitors from Medicinal Plants |
| title_sort | human tyrosinase displayed on the surface of chinese hamster ovary cells for ligand fishing of tyrosinase inhibitors from medicinal plants |
| topic | cell surface display Chinese hamster ovary cells tyrosinase inhibitors <i>Alhagi sparsifolia</i> <i>Coffea arabica</i> B16 melanoma cells |
| url | https://www.mdpi.com/1420-3049/30/1/30 |
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