Analysis of the mobility of signaling molecules in lymphocytes using fluorescence photobleaching techniques.
The earliest biochemical events at the plasma membrane that lead to gene activation appear to depend not only on the local concentration of signaling molecules, but also on the mobility of these molecules at the site of signaling. To elucidate the process of signal transduction after receptor engagement in the immune system, it is important to analyze the mobility of signaling molecules in living lymphocytes. Current knowledge of the changes in intracellular localization and dynamic movements of signaling molecules during lymphocyte activation is limited. Here, we describe a method for known as fluorescence recovery after photobleaching, used to measure the diffusion mobility of a signaling molecule in a T cell line after T cell receptor stimulation. This method is a useful tool in studies of spatiotemporal regulation in immunoreceptor signaling.
Tanimura
Natsuko
N
>Department of Immunobiology, Medical Technology, and Science, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan.
Nagafuku
Masakazu
M
Liddicoat
Douglas R
DR
Hamaoka
Toshiyuki
T
Kosugi
Atsushi
A
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D017951:Antigen Presentation; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D006224:Cricetinae; D036681:Fluorescence Recovery After Photobleaching / Q000295:instrumentation / Q000379:methods*; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins; D006801:Humans; D019169:Jurkat Cells; D008164:Luminescent Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology / Q000502:physiology; D008214:Lymphocytes / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D021962:Membrane Microdomains / Q000737:chemistry / Q000378:metabolism / Q000502:physiology; D008565:Membrane Proteins; D018613:Microscopy, Confocal; D008863:Microspheres; D016853:Muromonab-CD3 / Q000378:metabolism; D010750:Phosphoproteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D017260:Receptor-CD3 Complex, Antigen, T-Cell / Q000276:immunology / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology / Q000502:physiology*; D012984:Software; D013194:Staining and Labeling; D014162:Transfection
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; C110810:LAT protein, human; D008164:Luminescent Proteins; D008565:Membrane Proteins; D016853:Muromonab-CD3; D010750:Phosphoproteins; D017260:Receptor-CD3 Complex, Antigen, T-Cell; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins