Development and quantitative analysis of a biosensor based on the Arabidopsis SWEET1 sugar transporter
| Autor: | Jihyun Park, Taylor M. Chavez, Jordan A. Guistwhite, Sojeong Gwon, Wolf B. Frommer, Lily S. Cheung |
|---|---|
| Rok vydání: | 2021 |
| Předmět: | |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America |
| ISSN: | 1091-6490 |
| Popis: | Significance Transporters are the gatekeepers of the cell. Transporters facilitate the exchange of ions and metabolites between cellular and subcellular compartments, thus controlling processes from bacterial chemotaxis to the release of neurotransmitters. In plants, transporters have key roles in the allocation of carbon to nonphotosynthetic organs. Biosensors derived from transporters have been generated to monitor the activity of these proteins within the complex environment of the cell. However, a quantitative framework that reconciles molecular and cellular-level events to help interpret the response of biosensors is still lacking. Here, we created a sugar transporter biosensor and formulated a mathematical model to explain its response. These types of models can help realize multiscale, dynamic simulations of metabolite allocation to guide crop improvement. SWEETs are transporters with homologs in Archeae, plants, some fungi, and animals. As the only transporters known to facilitate the cellular release of sugars in plants, SWEETs play critical roles in the allocation of sugars from photosynthetic leaves to storage tissues in seeds, fruits, and tubers. Here, we report the design and use of genetically encoded biosensors to measure the activity of SWEETs. We created a SweetTrac1 sensor by inserting a circularly permutated green fluorescent protein into the Arabidopsis SWEET1, resulting in a chimera that translates substrate binding during the transport cycle into detectable changes in fluorescence intensity. We demonstrate that a combination of cell sorting and bioinformatics can accelerate the design of biosensors and formulate a mass action kinetics model to correlate the fluorescence response of SweetTrac1 with the transport of glucose. Our analysis suggests that SWEETs are low-affinity, symmetric transporters that can rapidly equilibrate intra- and extracellular concentrations of sugars. This approach can be extended to SWEET homologs and other transporters. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|