Quantitative analysis of confocal FRAP data
With the development of genetic means to labeling proteins in living cells with fluorescent proteins, it has become possible to monitor the kinetics of events such as protein diffusion in the cytoplasm or transient binding events to membranes using biophysical approaches such as confocal FRAP. Importantly, confocal FRAP is widely accessible to an exceptionally large number of experimentalists, and when analyzed using appropriate models, hold the potential to provide quantitative measurements of the on and off rates of binding, as well as directly measure the diffusion rate of the bound and unbound species. Work from my group in collaboration with bio-mathematicians has advanced knowledge in this arena through the development of mathematical models to analyze diffusion and reaction-diffusion events as tools to study intracellular protein and lipid dynamics.
Kang, M., C. A. Day, and A. K. Kenworthy (2019) A novel computational framework for D(t) from Fluorescence Recovery After Photobleaching data reveals various anomalous diffusion subtypes in live cell membranes. Traffic 20(11): 867-880.
Klaus, C.*, K. Raghunathan*, E. DiBenedetto, and A. K. Kenworthy (2016) Analysis of diffusion in curved surfaces and its application to tubular membranes. Mol Biol Cell 27(24): 3937-3946.DOI: 10.1091/mbc.E16-06-0445. PMCID: PMC5170615. *equal contributions
Kang, M. Andreani, M., and A. K. Kenworthy (2015) Validation of normalizations, scaling, and photo fading corrections for FRAP data analysis. PLoS ONE 10(5): e0127966
Kang, M., C. A. Day, A. K. Kenworthy and E. DiBenedetto (2012) Simplified equation to extract diffusion coefficients from confocal FRAP data. Traffic 13(12): 1589-600. PMCID: PMC3731631
Day. C. A., L. J. Kraft, M. Kang, and A. K. Kenworthy (2012) Analysis of protein and lipid dynamics using confocal fluorescence recovery after photobleaching. Current Protocols in Cytometry, Chapter 2: Unit 2.19.PMCID: PMC3538152
Kang, M., C. A. Day, E.DiBenedetto and A. K. Kenworthy (2010) A quantitative approach to analyze binding diffusion kinetics by confocal FRAP. Biophys J 99(9): 2737-47. PMCID: PMC2965996
Kang, M., C. A. Day, K. Drake, A. K. Kenworthy and E. DiBenedetto (2009) A generalization of theory for two-dimensional fluorescence recovery after photobleaching applicable to confocal laser scanning microscopes. Biophys J 97(5): 1501-11. PMCID: PMC2749741
Kang, M. and A. K. Kenworthy (2008) A closed-form analytic expression for FRAP formula for the binding diffusion model. Biophys J 95(2): L13-5. PMCID: PMC2440465
Goodwin, J. S., K. R. Drake, C. Rogers, L. Wright, J. Lippincott-Schwartz, M. R. Philips and A. K. Kenworthy (2005). Depalmitoylated Ras traffics to and from the Golgi complex via a nonvesicular pathway. J Cell Biol 170(2): 261-72.