Anti-nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane protein
Abstract Nanodisc technology has dramatically advanced the analysis of molecular interactions for membrane proteins. A nanodisc is designed as a vehicle for membrane proteins that provide a native-like phospholipid environment and better thermostability in a detergent-free buffer. This enables the d...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-07-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-38547-2 |
| _version_ | 1797774270535827456 |
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| author | Fuhito Nakagawa Marin Kikkawa Sisi Chen Yasuomi Miyashita Norie Hamaguchi-Suzuki Minami Shibuya Soichi Yamashita Lisa Nagase Satoshi Yasuda Mitsunori Shiroishi Toshiya Senda Keisuke Ito Takeshi Murata Satoshi Ogasawara |
| author_facet | Fuhito Nakagawa Marin Kikkawa Sisi Chen Yasuomi Miyashita Norie Hamaguchi-Suzuki Minami Shibuya Soichi Yamashita Lisa Nagase Satoshi Yasuda Mitsunori Shiroishi Toshiya Senda Keisuke Ito Takeshi Murata Satoshi Ogasawara |
| author_sort | Fuhito Nakagawa |
| collection | DOAJ |
| description | Abstract Nanodisc technology has dramatically advanced the analysis of molecular interactions for membrane proteins. A nanodisc is designed as a vehicle for membrane proteins that provide a native-like phospholipid environment and better thermostability in a detergent-free buffer. This enables the determination of the thermodynamic and kinetic parameters of small molecule binding by surface plasmon resonance. In this study, we generated a nanodisc specific anti-MSP (membrane scaffold protein) monoclonal antibody biND5 for molecular interaction analysis of nanodiscs. The antibody, biND5 bound to various types of nanodiscs with sub-nanomolar to nanomolar affinity. Epitope mapping analysis revealed specific recognition of 8 amino acid residues in the exposed helix-4 structure of MSP. Further, we performed kinetics binding analysis between adenosine A2a receptor reconstituted nanodiscs and small molecule antagonist ZM241385 using biND5 immobilized sensor chips. These results show that biND5 facilitates the molecular interaction kinetics analysis of membrane proteins substituted in nanodiscs. |
| first_indexed | 2024-03-12T22:17:39Z |
| format | Article |
| id | doaj.art-311e47254bf243f790c71dd2fbab2556 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-03-12T22:17:39Z |
| publishDate | 2023-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-311e47254bf243f790c71dd2fbab25562023-07-23T11:12:15ZengNature PortfolioScientific Reports2045-23222023-07-0113111110.1038/s41598-023-38547-2Anti-nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane proteinFuhito Nakagawa0Marin Kikkawa1Sisi Chen2Yasuomi Miyashita3Norie Hamaguchi-Suzuki4Minami Shibuya5Soichi Yamashita6Lisa Nagase7Satoshi Yasuda8Mitsunori Shiroishi9Toshiya Senda10Keisuke Ito11Takeshi Murata12Satoshi Ogasawara13Department of Chemistry, Graduate School of Science, Chiba UniversityDepartment of Food and Nutritional Sciences, Graduate School of Integrated Pharmaceutical and Nutritional Sciences, University of ShizuokaDepartment of Chemistry, Graduate School of Science, Chiba UniversityDepartment of Chemistry, Graduate School of Science, Chiba UniversityDepartment of Chemistry, Graduate School of Science, Chiba UniversityDepartment of Chemistry, Graduate School of Science, Chiba UniversityDepartment of Chemistry, Graduate School of Science, Chiba UniversityStructure Biology Research Center, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK)Department of Chemistry, Graduate School of Science, Chiba UniversityDepartment of Biological Science and Technology, Tokyo University of ScienceStructure Biology Research Center, Institute of Materials Structure Science, High Energy Accelerator Research Organization (KEK)Department of Food and Nutritional Sciences, Graduate School of Integrated Pharmaceutical and Nutritional Sciences, University of ShizuokaDepartment of Chemistry, Graduate School of Science, Chiba UniversityDepartment of Chemistry, Graduate School of Science, Chiba UniversityAbstract Nanodisc technology has dramatically advanced the analysis of molecular interactions for membrane proteins. A nanodisc is designed as a vehicle for membrane proteins that provide a native-like phospholipid environment and better thermostability in a detergent-free buffer. This enables the determination of the thermodynamic and kinetic parameters of small molecule binding by surface plasmon resonance. In this study, we generated a nanodisc specific anti-MSP (membrane scaffold protein) monoclonal antibody biND5 for molecular interaction analysis of nanodiscs. The antibody, biND5 bound to various types of nanodiscs with sub-nanomolar to nanomolar affinity. Epitope mapping analysis revealed specific recognition of 8 amino acid residues in the exposed helix-4 structure of MSP. Further, we performed kinetics binding analysis between adenosine A2a receptor reconstituted nanodiscs and small molecule antagonist ZM241385 using biND5 immobilized sensor chips. These results show that biND5 facilitates the molecular interaction kinetics analysis of membrane proteins substituted in nanodiscs.https://doi.org/10.1038/s41598-023-38547-2 |
| spellingShingle | Fuhito Nakagawa Marin Kikkawa Sisi Chen Yasuomi Miyashita Norie Hamaguchi-Suzuki Minami Shibuya Soichi Yamashita Lisa Nagase Satoshi Yasuda Mitsunori Shiroishi Toshiya Senda Keisuke Ito Takeshi Murata Satoshi Ogasawara Anti-nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane protein Scientific Reports |
| title | Anti-nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane protein |
| title_full | Anti-nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane protein |
| title_fullStr | Anti-nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane protein |
| title_full_unstemmed | Anti-nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane protein |
| title_short | Anti-nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane protein |
| title_sort | anti nanodisc antibodies specifically capture nanodiscs and facilitate molecular interaction kinetics studies for membrane protein |
| url | https://doi.org/10.1038/s41598-023-38547-2 |
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