NR AVFB
AU Wu,Y.X.; Masison,D.C.; Eisenberg,E.; Greene,L.E.
TI Application of photobleaching for measuring diffusion of prion proteins in cytosol of yeast cells
QU Methods (San Diego, Calif.) 2006 May; 39(1): 43-9
PT journal article
AB Measurement of fluorescence recovery after photobleaching (FRAP) is a non-invasive technique for studying protein dynamics in real time in living cells. FRAP studies are carried out on proteins tagged with green fluorescent protein (GFP) or one of its spectral variants. Illumination with high intensity laser light irreversibly bleaches the GFP fluorescence but has no effect on protein function. By photobleaching a limited region of the cytoplasm, the rate of fluorescence recovery provides a measure of the rate of protein diffusion. A detailed description of the FRAP technique is given, including its application to measuring the mobility of GFP-tagged Sup35p in [psi(-)] and [PSI(+)] cells.
MH Algorithms; Cytosol/*metabolism; Diffusion; Fluorescence Recovery After Photobleaching/*methods; Green Fluorescent Proteins/chemistry/genetics/metabolism; Prions/genetics/*metabolism; Recombinant Fusion Proteins/chemistry/genetics/metabolism; Saccharomyces cerevisiae/genetics/*metabolism; Saccharomyces cerevisiae Proteins/genetics/*metabolism
AD Laboratory of Cell Biology, NHLBI, NIH, Bethesda, MD 20892-0301, USA
SP englisch
PO USA