Membrane bound protein diffusion viewed by fluorescence recovery after bleaching experiments : models analysis
نویسندگان
چکیده
Membrane bound protein diffusion viewed by fluorescence recovery after bleaching experiments : models analysis. Abstract Diffusion processes in biological membranes are of interest to understand the macromolecular organisation and function of several molecules. Fluorescence Recovery After Photobleaching (FRAP) has been widely used as a method to analyse this processes using classical Brownian diffusion model. In the first part of this work, the analytical expression of the fluorescence recovery as a function of time has been established for anomalous diffusion due to long waiting times. Then, experimental fluorescence recoveries recorded in living cells on a membrane-bound protein have been analysed using three different models : normal Brownian diffusion, Brownian diffusion with an immobile fraction and anomalous diffusion due to long waiting times.
منابع مشابه
Spontaneous recovery of fluorescence by photobleached surface-adsorbed proteins.
Fluorescence photobleaching of a carboxyfluorescein-labeled protein (erythrocyte cytoskeletal protein 4.1) immobilized on bare glass is found to be spontaneously reversible, provided that the sample is deoxygenated. After a short (hundredths of seconds) photobleaching laser flash, the subsequent fluorescence excited by a dim probe beam partly recovers on a long (tenths of second) time scale, ev...
متن کاملFRAP to Characterize Molecular Diffusion and Interaction in Various Membrane Environments
Fluorescence recovery after photobleaching (FRAP) is a standard method used to study the dynamics of lipids and proteins in artificial and cellular membrane systems. The advent of confocal microscopy two decades ago has made quantitative FRAP easily available to most laboratories. Usually, a single bleaching pattern/area is used and the corresponding recovery time is assumed to directly provide...
متن کاملFluorescence recovery after photo-bleaching as a method to determine local diffusion coefficient in the stratum corneum.
Fluorescence recovery after photo-bleaching experiments were performed in human stratum corneum in vitro. Fluorescence multiphoton tomography was used, which allowed the dimensions of the photobleached volume to be at the micron scale and located fully within the lipid phase of the stratum corneum. Analysis of the fluorescence recovery data with simplified mathematical models yielded the diffus...
متن کاملRestricted mobility of membrane constituents in cell-substrate focal contacts of chicken fibroblasts
We studied the lateral mobility of membrane components in cell-substrate focal contacts using the fluorescence photobleaching recovery method. The measurements were performed on isolated substrate-attached membranes of chicken gizzard fibroblasts. The diffusion coefficients of a fluorescent lipid probe and rhodamine-conjugated surface proteins within contact regions (identified by interference-...
متن کاملPerformance Evaluation of Ceramic Membrane on Ultrafiltration and Diafiltration Modes for Efficient Recovery of Whey Protein
The recovery of high-quality, valuable end products from cheese whey is a very important industrial process. The aim of this study is to improve the recovered protein content of concentrate stream in cheese whey. The experiments were conducted by using ceramic ultrafiltration (UF) membrane followed by diafiltration (DF) process with a 15kDa molecular weight cut-off (MWCO). The volume concentrat...
متن کامل