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Constitutive plasma membrane targeting and microdomain localization of Dok5 studied by single-molecule microscopy
Authors:Fu Guo  Zhang Feng  Cao Li  Xu Zhi-Zhan  Chen Yi-Zhang  Wang Gui-Ying  He Cheng
Institution:Institute of Neuroscience, Department of Neurobiology, Second Military Medical University, Shanghai 200433, China.
Abstract:In the present study, single-molecule fluorescence microscopy was used to examine the characteristics of plasma membrane targeting and microdomain localization of enhanced yellow fluorescent protein (eYFP)-tagged wild-type Dok5 and its variants in living Chinese hamster ovary (CHO) cells. We found that Dok5 can target constitutively to the plasma membrane, and the PH domain is essential for this process. Furthermore, single-molecule trajectories analysis revealed that Dok5 can constitutively partition into microdomain on the plasma membrane. Finally, the potential mechanism of microdomain localization of Dok5 was discussed. This study provided insights into the characteristics of plasma membrane targeting and microdomain localization of Dok5 in living CHO cells.
Keywords:Dok  Single-molecule  Pleckstrin homology domain  Microdomain  Diffusion  Total internal reflection fluorescence microscopy
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