ArticlePhysical biology2010
Analysis of the diffusion of Ras2 in Saccharomyces cerevisiae using fluorescence recovery after photobleaching.
Article in Physical biology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed.
- Evaluation of Lipids and Lipid-Related Transcripts in Human and Ovine Theca Cells and anEnvironmental health perspectives · 2024Article
- Multiscale model of the physiological control of myocardial perfusion to delineate putative metabolic feedback mechanisms.The Journal of physiology · 2022Article
- Membrane thickness, lipid phase and sterol type are determining factors in the permeability of membranes to small solutes.Nature communications · 2022Article
- Cell patterning by secretion-induced plasma membrane flows.Science advances · 2021Article
- DeepFRAP: Fast fluorescence recovery after photobleaching data analysis using deep neural networks.Journal of microscopy · 2021Article
- A Highly Accurate Pixel-Based FRAP Model Based on Spectral-Domain Numerical Methods.Biophysical journal · 2019Article
- Exploration and stabilization of Ras1 mating zone: A mechanism with positive and negative feedbacks.PLoS computational biology · 2018Article
- Characterization of Cell Boundary and Confocal Effects Improves Quantitative FRAP Analysis.Biophysical journal · 2018Article
- Steric exclusion and protein conformation determine the localization of plasma membrane transporters.Nature communications · 2018Article
- Roles of Ras1 membrane localization during Candida albicans hyphal growth and farnesol response.Eukaryotic cell · 2011Article
- Phosphatidylserine is polarized and required for proper Cdc42 localization and for development of cell polarity.Nature cell biology · 2011Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Binding, lateral diffusion and exchange are fundamental dynamic processes involved in protein association with cellular membranes. In this study, we developed numerical simulations of lateral diffusion and exchange of fluorophores in membranes with arbitrary bleach geometry and exchange of the membrane-localized fluorophore with the cytosol during fluorescence recovery after photobleaching (FRAP) experiments. The model simulations were used to design FRAP experiments with varying bleach region sizes on plasma membrane-localized wild-type GFP-Ras2 with a dual lipid anchor and mutant GFP-Ras2C318S with a single lipid anchor in live yeast cells to investigate diffusional mobility and the presence of any exchange processes operating in the time scale of our experiments. Model parameters estimated using data from FRAP experiments with a 1 microm x 1 microm bleach region-of-interest (ROI) and a 0.5 microm x 0.5 microm bleach ROI showed that GFP-Ras2, single or dual lipid modified, diffuses as single species with no evidence of exchange with a cytoplasmic pool. This is the first report of Ras2 mobility in the yeast plasma membrane. The methods developed in this study are generally applicable for studying diffusion and exchange of membrane-associated fluorophores using FRAP on commercial confocal laser scanning microscopes.
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