ArticleBiophysical journal2010
A quantitative approach to analyze binding diffusion kinetics by confocal FRAP.
Article in Biophysical journal, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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Who cites it
32 citing papers in PubMed.
- Inferring diffusion, reaction, and exchange parameters from imperfect FRAP.Biophysical journal · 2025Article
- Transport Properties of Self-Assembling G-Hydrogels: Evidence for a Tunable Fickian Diffusivity.The journal of physical chemistry. B · 2025Article
- Advantages and Limitations of Photoconvertible Probes to Study Subcellular Dynamics in Epithelial Cells.Biology of the cell · 2025Article
- Article
- Beyond analytic solution: Analysis of FRAP experiments by spatial simulation of the forward problem.Biophysical journal · 2023Article
- What's past is prologue: FRAP keeps delivering 50 years later.Biophysical journal · 2023Review
- Use of Fluorescence Recovery After Photobleaching (FRAP) to Measure In Vivo Dynamics of Cell Junction-Associated Polarity Proteins.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Article
- Parameter estimation in fluorescence recovery after photobleaching: quantitative analysis of protein binding reactions and diffusion.Journal of mathematical biology · 2021Article
- DeepFRAP: Fast fluorescence recovery after photobleaching data analysis using deep neural networks.Journal of microscopy · 2021Article
- Self-sorting in supramolecular assemblies.Soft matter · 2021Article
- A Simple Derivation of Diffusion Fluorescence Correlation Spectroscopy Equations.Journal of fluorescence · 2020Article
- Article
- Bound-State Diffusion due to Binding to Flexible Polymers in a Selective Biofilter.Biophysical journal · 2020Article
- A Highly Accurate Pixel-Based FRAP Model Based on Spectral-Domain Numerical Methods.Biophysical journal · 2019Article
- A micropatterning platform for quantifying interaction kinetics between the T cell receptor and an intracellular binding protein.Scientific reports · 2019Article
- The Development and Enhancement of FRAP as a Key Tool for Investigating Protein Dynamics.Biophysical journal · 2018Review
- Quantitative Analysis Reveals that Actin and Src-Family Kinases Regulate Nuclear YAP1 and Its Export.Cell systems · 2018Article
- Characterization of Cell Boundary and Confocal Effects Improves Quantitative FRAP Analysis.Biophysical journal · 2018Article
- Analysis of Active Transport by Fluorescence Recovery after Photobleaching.Biophysical journal · 2017Article
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Authors and funding
4 authors.
Funding
Abstract
Most of the important types of interactions that occur in cells can be characterized as binding-diffusion type processes, and can be quantified by kinetic rate constants such as diffusion coefficients (D) and binding rate constants (k(on) and k(off)). Confocal FRAP is a potentially important tool for the quantitative analysis of intracellular binding-diffusion kinetics, but how to dependably extract accurate kinetic constants from such analyses is still an open question. To this end, in this study, we developed what we believe is a new analytical model for confocal FRAP-based measurements of intracellular binding-diffusion processes, based on a closed-form equation of the FRAP formula for a spot photobleach geometry. This approach incorporates a binding diffusion model that allows for diffusion of both the unbound and bound species, and also compensates for binding diffusion that occurs during photobleaching, a critical consideration in confocal FRAP analysis. In addition, to address the problem of parametric multiplicity, we propose a scheme to reduce the number of fitting parameters in the effective diffusion subregime when D's for the bound and unbound species are known. We validate this method by measuring kinetic rate constants for the CAAX-mediated binding of Ras to membranes of the endoplasmic reticulum, obtaining binding constants of k(on) ∼ 255/s and k(off) ∼ 31/s.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.