Protocol for measuring D-amino acid transport in mammalian cells using a fluorescent biosensor
Summary: The movement of amino acids into cells and organelles is facilitated by transporter proteins, which support homeostasis, metabolism, and disease progression. Here, we present a protocol to visualize amino acid transport in mammalian cells in real time using an enzyme-based biosensor. We pro...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725003351 |
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| author | Danisha Johal Keeley G. Hewton Sadie J. Gale Amritpal S. Johal Seth J. Parker |
| author_facet | Danisha Johal Keeley G. Hewton Sadie J. Gale Amritpal S. Johal Seth J. Parker |
| author_sort | Danisha Johal |
| collection | DOAJ |
| description | Summary: The movement of amino acids into cells and organelles is facilitated by transporter proteins, which support homeostasis, metabolism, and disease progression. Here, we present a protocol to visualize amino acid transport in mammalian cells in real time using an enzyme-based biosensor. We provide instructions for plating sensor-expressing HEK293 cells, optimizing fluorescent readouts, and initiating transport measurements. Data interpretation and analysis are also discussed.For complete details on the use and execution of this protocol, please refer to Johal et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-6b0147cef77d4696936e8dc0b86710e5 |
| institution | Matheson Library |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-6b0147cef77d4696936e8dc0b86710e52025-06-30T04:09:15ZengElsevierSTAR Protocols2666-16672025-09-0163103929Protocol for measuring D-amino acid transport in mammalian cells using a fluorescent biosensorDanisha Johal0Keeley G. Hewton1Sadie J. Gale2Amritpal S. Johal3Seth J. Parker4Department of Biochemistry & Molecular Biology, University of British Columbia, Vancouver, BC V6T 1Z4, Canada; Corresponding authorDepartment of Biochemistry & Molecular Biology, University of British Columbia, Vancouver, BC V6T 1Z4, Canada; Corresponding authorDepartment of Biochemistry & Molecular Biology, University of British Columbia, Vancouver, BC V6T 1Z4, CanadaDepartment of Biochemistry & Molecular Biology, University of British Columbia, Vancouver, BC V6T 1Z4, CanadaDepartment of Biochemistry & Molecular Biology, University of British Columbia, Vancouver, BC V6T 1Z4, Canada; Centre for Molecular Medicine and Therapeutics, University of British Columbia, Vancouver, BC V5Z 4H4, Canada; British Columbia Children’s Hospital Research Institute, Vancouver, BC V5Z 4H4, Canada; Corresponding authorSummary: The movement of amino acids into cells and organelles is facilitated by transporter proteins, which support homeostasis, metabolism, and disease progression. Here, we present a protocol to visualize amino acid transport in mammalian cells in real time using an enzyme-based biosensor. We provide instructions for plating sensor-expressing HEK293 cells, optimizing fluorescent readouts, and initiating transport measurements. Data interpretation and analysis are also discussed.For complete details on the use and execution of this protocol, please refer to Johal et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725003351Cell MembraneCancerMetabolismMolecular/Chemical Probes |
| spellingShingle | Danisha Johal Keeley G. Hewton Sadie J. Gale Amritpal S. Johal Seth J. Parker Protocol for measuring D-amino acid transport in mammalian cells using a fluorescent biosensor STAR Protocols Cell Membrane Cancer Metabolism Molecular/Chemical Probes |
| title | Protocol for measuring D-amino acid transport in mammalian cells using a fluorescent biosensor |
| title_full | Protocol for measuring D-amino acid transport in mammalian cells using a fluorescent biosensor |
| title_fullStr | Protocol for measuring D-amino acid transport in mammalian cells using a fluorescent biosensor |
| title_full_unstemmed | Protocol for measuring D-amino acid transport in mammalian cells using a fluorescent biosensor |
| title_short | Protocol for measuring D-amino acid transport in mammalian cells using a fluorescent biosensor |
| title_sort | protocol for measuring d amino acid transport in mammalian cells using a fluorescent biosensor |
| topic | Cell Membrane Cancer Metabolism Molecular/Chemical Probes |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725003351 |
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