From fixed to FRAP: measuring protein mobility and activity in living cells

被引:490
作者
Reits, EAJ [1 ]
Neefjes, JJ [1 ]
机构
[1] Netherlands Canc Inst, Div Tumor Biol, NL-1066 CX Amsterdam, Netherlands
关键词
D O I
10.1038/35078615
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Experiments with fluorescence recovery after photobleaching (FRAP) started 30 years ago to visualize the lateral mobility and dynamics of fluorescent proteins in living cells. Its popularity increased when non-invasive fluorescent tagging became possible with the green fluorescent protein (GFP). Many researchers use GFP to study the localization of fusion proteins in fixed or living cells, but the same fluorescent proteins can also be used to study protein mobility in living cells. Here we review the potential of FRAP to study protein dynamics and activity within a single living cell. These measurements can be made with most standard confocal laser-scanning microscopes equipped with photobleaching protocols.
引用
收藏
页码:E145 / E147
页数:3
相关论文
共 29 条
[1]   USE OF A FLUORESCENT CHOLESTEROL DERIVATIVE TO MEASURE LATERAL MOBILITY OF CHOLESTEROL IN MEMBRANES [J].
ALECIO, MR ;
GOLAN, DE ;
VEATCH, WR ;
RANDO, RR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1982, 79 (17) :5171-5174
[2]   Mobility of creatine phosphokinase and beta-enolase in cultured muscle cells [J].
ArrioDupont, M ;
Foucault, G ;
Vacher, M ;
Douhou, A ;
Cribier, S .
BIOPHYSICAL JOURNAL, 1997, 73 (05) :2667-2673
[3]   Diffusion of fluorescently labeled macromolecules in cultured muscle cells [J].
ArrioDupont, M ;
Cribier, S ;
Foucault, G ;
Devaux, PF ;
dAlbis, A .
BIOPHYSICAL JOURNAL, 1996, 70 (05) :2327-2332
[4]   MOBILITY MEASUREMENT BY ANALYSIS OF FLUORESCENCE PHOTOBLEACHING RECOVERY KINETICS [J].
AXELROD, D ;
KOPPEL, DE ;
SCHLESSINGER, J ;
ELSON, E ;
WEBB, WW .
BIOPHYSICAL JOURNAL, 1976, 16 (09) :1055-1069
[5]   LATERAL MOTION OF FLUORESCENTLY LABELED ACETYLCHOLINE RECEPTORS IN MEMBRANES OF DEVELOPING MUSCLE-FIBERS [J].
AXELROD, D ;
RAVDIN, P ;
KOPPEL, DE ;
SCHLESSINGER, J ;
WEBB, WW ;
ELSON, EL ;
PODLESKI, TR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1976, 73 (12) :4594-4598
[6]   Diffusional mobility of Golgi proteins in membranes of living cells [J].
Cole, NB ;
Smith, CL ;
Sciaky, N ;
Terasaki, M ;
Edidin, M ;
LippincottSchwartz, J .
SCIENCE, 1996, 273 (5276) :797-801
[7]   Diffusion of green fluorescent protein in the aqueous-phase lumen of endoplasmic reticulum [J].
Dayel, MJ ;
Hom, EFY ;
Verkman, AS .
BIOPHYSICAL JOURNAL, 1999, 76 (05) :2843-2851
[8]   The dynamics of postmitotic reassembly of the nucleolus [J].
Dundr, M ;
Misteli, T ;
Olson, MOJ .
JOURNAL OF CELL BIOLOGY, 2000, 150 (03) :433-446
[9]   TRUNCATION MUTANTS DEFINE AND LOCATE CYTOPLASMIC BARRIERS TO LATERAL MOBILITY OF MEMBRANE-GLYCOPROTEINS [J].
EDIDIN, M ;
ZUNIGA, MC ;
SHEETZ, MP .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (08) :3378-3382
[10]  
EDIDIN M, 1976, SCIENCE, V191, P466, DOI 10.1126/science.1246629