-
Real-time detection of changes in yeast plasma membrane potential using genetically encoded voltage indicator proteins
- Back
Document Title
Real-time detection of changes in yeast plasma membrane potential using genetically encoded voltage indicator proteins
Author
Limapichat W., Pornthanakasem W., Satitthammachart C., Chitnumsub P., Leartsakulpanich U.
Name from Authors Collection
Affiliations
National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park, Phahonyothin Road, Khlong Nueng, Khlong Luang, Pathum Thani, 12120, Thailand
Type
Article
Source Title
FEMS Yeast Research
ISSN
15671356
Year
2020
Volume
20
Issue
5
Open Access
All Open Access, Bronze
Publisher
Oxford University Press
DOI
10.1093/femsyr/foaa041
Format
Abstract
In yeast, adaptation to varying conditions often requires proper regulation of the plasma membrane potential. To determine yeast membrane potential change, optical methods involving potentiometric dyes have been supplemental to the direct electrode-based method. However, the hydrophobic nature of the dyes and their slow distribution across the membrane still limits their utilization. Genetically encoded voltage indicator (GEVI) proteins employed in neuroscience offer a tantalizing alternative for monitoring yeast membrane potential change. In this work, several widely used GEVI proteins were assessed in Saccharomyces cerevisiae for their expression and function as a voltage reporter. Among them, only ArcLight and Accelerated Sensor of Action Potential (ASAP) proteins could be expressed and transported to the plasma membrane. While the voltage-sensing capability was demonstrated for both ArcLight and ASAP, ArcLight fluorescence was sensitive to the intracellular pH change concurrently with the voltage change. Therefore, we established that ASAP is the more suitable GEVI protein for reporting yeast membrane potential change. This voltage-sensing reporter for yeast based on ASAP offers a new effective strategy for real-time optical detection of yeast membrane potential change, which potentially facilitates many areas of yeast research including optimizing growth conditions for industrial use and investigating yeast ion transport system. © FEMS 2020. All rights reserved.
Industrial Classification
Knowledge Taxonomy Level 1
Knowledge Taxonomy Level 2
Knowledge Taxonomy Level 3
Funding Sponsor
National Science and Technology Development Agency; Thailand Research Fund
Funding Number
P16-50382, P18-51412, P18-51598
License
N/A
Rights
N/A
Publication Source
Scopus