By Yasushi Sako, Masahiro Ueda
This booklet encompasses the intriguing advancements and demanding situations within the fast-moving and swiftly increasing study box of single-molecule kinetic research of phone signaling that provides to be some of the most major and fascinating parts of organic learn for the foreseeable destiny. cellphone signaling is conducted by means of advanced response networks of macromolecules, and single-molecule analyses has already confirmed its energy to solve complicated response dynamics in purified platforms. so far, many of the released study within the box of single-molecule tactics in cells, specialize in the dynamic homes (translational events of the centre of mass) of organic molecules. in spite of the fact that, we are hoping that this e-book provides as many kinetic analyses of cellphone signaling as attainable. even if single-molecule kinetic research of mobile platforms is a comparatively younger box compared to the research of single-molecule hobbies in cells, this sort of research is extremely very important since it at once pertains to the molecular capabilities that keep watch over mobile habit and sooner or later, single-molecule kinetic research can be mostly directed in the direction of mobile structures. therefore, we are hoping that this e-book might be of curiosity to all these operating within the fields of molecular and mobile biology, in addition to biophysics and biochemistry.
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Extra info for Cell Signaling Reactions: Single-Molecular Kinetic Analysis
Teramura Y, Ichinose J, Takagi H, Nishida K, Yanagida T, Sako Y (2006) Single molecule analysis of epidermal growth factor binding on the surface of living cells. EMBO J 25:4215 4222 83. Tokunaga M, Imamoto N, Sakata Sogawa K (2008) Highly inclined thin illumination enables clear single molecule imaging in cells. Nat Methods 5:159 161 84. Tokunaga M, Kitamura K, Saito K, Iwane AH, Yanagida T (1997) Single molecule imaging of fluorophores enzymatic reactions achieved by objective type total internal reflection fluorescence microscopy.
Webb SED, Roberts SK, Needham SR, Tynan CJ, Rolfe DJ, Winn MD, Clarke DT, Barraclough R, Martin Fernandez ML (2008) Single molecule imaging and fluorescence 32 91. 92. 93. 94. 95. 96. 97. M. Hiroshima and Y. Sako lifetime imaging microscopy show different structures for high and low affinity epidermal growth factor receptors in A431 cells. Biophys J 94:803 819 Xiao Z, Ma X, Jiang Y, Zhao Z, Lai B, Liao J, Yue J, Fang X (2008) Single molecule study of lateral mobility of epidermal growth factor receptor 2/HER2 on activation.
The intracellular Ca2+ response is preceded by EGF-induced receptor dimerization. We tried to clarify the 20 M. Hiroshima and Y. Sako Percentage of positive cells quantitative relationship between the EGF binding to EGFR and the Ca2+ response using single-molecule techniques. In A431 cells, single-molecule quantification of receptor-bound Cy3-EGF and measurement of the signal intensity of the Ca2+ indicator Fluo-3 in the cytoplasm showed that EGF dimers increased for 60 s after the application of Cy3-EGF to the extracellular medium and that the Ca2+ concentration subsequently increased .