Optical Bioimaging: From Living Tissue to a Single Molecule: Single-Molecule Visualization of Cell Signaling Processes of Epidermal Growth Factor Receptor
ABSTRACT: Single-molecule imaging is an ideal technology to study molecular mechanisms of biological reactions in vitro. Recently, this technology has been extended to real-time observation of fluorescent dye-labeled molecules in living cells. Total internal reflection fluorescence microscopy is the...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2003-01-01
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| Series: | Journal of Pharmacological Sciences |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1347861319325733 |
| _version_ | 1819092196721164288 |
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| author | Yasushi Sako Junya Ichinose Miki Morimatsu Kosuke Ohta Takeshi Uyemura |
| author_facet | Yasushi Sako Junya Ichinose Miki Morimatsu Kosuke Ohta Takeshi Uyemura |
| author_sort | Yasushi Sako |
| collection | DOAJ |
| description | ABSTRACT: Single-molecule imaging is an ideal technology to study molecular mechanisms of biological reactions in vitro. Recently, this technology has been extended to real-time observation of fluorescent dye-labeled molecules in living cells. Total internal reflection fluorescence microscopy is the major technique for this purpose. Using this technique, we have studied the process of early signal transduction of epidermal growth factor (EGF) in single molecules: binding of EGF to its receptor (EGFR) on the cell surface, dimerization of EGFR induced by binding of EGF, fluctuation of the structure of EGFR clusters, activation of EGFR through tyrosine phosphorylations on its cytoplasmic domain, and recognition of activated EGFR by a cytoplasmic adaptor protein, Grb2. EGF induces intracellular calcium response, sometimes caused by less than one hundred EGF molecules. Single-molecule studies suggested that this highly sensitive response to EGF was due to the amplification of the EGFR signal using dynamic clustering, reorganization of the dimers, and lateral mobility of EGFR on the cell surface. Through these studies, single-molecule analysis has proven to be a powerful technology to analyze intracellular protein systems. |
| first_indexed | 2024-12-21T22:51:47Z |
| format | Article |
| id | doaj.art-a1b4f558fad74fb999edafd5ef02e7c3 |
| institution | Directory Open Access Journal |
| issn | 1347-8613 |
| language | English |
| last_indexed | 2024-12-21T22:51:47Z |
| publishDate | 2003-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Pharmacological Sciences |
| spelling | doaj.art-a1b4f558fad74fb999edafd5ef02e7c32022-12-21T18:47:34ZengElsevierJournal of Pharmacological Sciences1347-86132003-01-01933253258Optical Bioimaging: From Living Tissue to a Single Molecule: Single-Molecule Visualization of Cell Signaling Processes of Epidermal Growth Factor ReceptorYasushi Sako0Junya Ichinose1Miki Morimatsu2Kosuke Ohta3Takeshi Uyemura4Nanobiology Laboratories, Graduate School of Frontier Biosciences, Osaka University, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan; Time’s Arrow and Biosignaling, PRESTO, JSTTime’s Arrow and Biosignaling, PRESTO, JSTTime’s Arrow and Biosignaling, PRESTO, JSTDepartment of Systems and Human Science, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama Toyonaka, Osaka 560-8531, JapanDepartment of Systems and Human Science, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama Toyonaka, Osaka 560-8531, JapanABSTRACT: Single-molecule imaging is an ideal technology to study molecular mechanisms of biological reactions in vitro. Recently, this technology has been extended to real-time observation of fluorescent dye-labeled molecules in living cells. Total internal reflection fluorescence microscopy is the major technique for this purpose. Using this technique, we have studied the process of early signal transduction of epidermal growth factor (EGF) in single molecules: binding of EGF to its receptor (EGFR) on the cell surface, dimerization of EGFR induced by binding of EGF, fluctuation of the structure of EGFR clusters, activation of EGFR through tyrosine phosphorylations on its cytoplasmic domain, and recognition of activated EGFR by a cytoplasmic adaptor protein, Grb2. EGF induces intracellular calcium response, sometimes caused by less than one hundred EGF molecules. Single-molecule studies suggested that this highly sensitive response to EGF was due to the amplification of the EGFR signal using dynamic clustering, reorganization of the dimers, and lateral mobility of EGFR on the cell surface. Through these studies, single-molecule analysis has proven to be a powerful technology to analyze intracellular protein systems.http://www.sciencedirect.com/science/article/pii/S1347861319325733 |
| spellingShingle | Yasushi Sako Junya Ichinose Miki Morimatsu Kosuke Ohta Takeshi Uyemura Optical Bioimaging: From Living Tissue to a Single Molecule: Single-Molecule Visualization of Cell Signaling Processes of Epidermal Growth Factor Receptor Journal of Pharmacological Sciences |
| title | Optical Bioimaging: From Living Tissue to a Single Molecule: Single-Molecule Visualization of Cell Signaling Processes of Epidermal Growth Factor Receptor |
| title_full | Optical Bioimaging: From Living Tissue to a Single Molecule: Single-Molecule Visualization of Cell Signaling Processes of Epidermal Growth Factor Receptor |
| title_fullStr | Optical Bioimaging: From Living Tissue to a Single Molecule: Single-Molecule Visualization of Cell Signaling Processes of Epidermal Growth Factor Receptor |
| title_full_unstemmed | Optical Bioimaging: From Living Tissue to a Single Molecule: Single-Molecule Visualization of Cell Signaling Processes of Epidermal Growth Factor Receptor |
| title_short | Optical Bioimaging: From Living Tissue to a Single Molecule: Single-Molecule Visualization of Cell Signaling Processes of Epidermal Growth Factor Receptor |
| title_sort | optical bioimaging from living tissue to a single molecule single molecule visualization of cell signaling processes of epidermal growth factor receptor |
| url | http://www.sciencedirect.com/science/article/pii/S1347861319325733 |
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