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KMID : 0613820220320020148
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Journal of Life Science
2022 Volume.32 No. 2 p.148 ~ p.154
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Single-Cell-Imaging-Based Analysis of Focal Adhesion Kinase Activity in Plasma Membrane Microdomains Under a Diverse Composition of Extracellular Matrix Proteins
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Choi Gyu-Ho
Jang Yoon-Kwan
Suh Jung-Soo
Kim Heon-Su
Ahn Sang-Hyun
Han Ki-Seok
Kim Eun-Hye
Kim Tae-Jin
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Abstract
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Focal adhesion kinase (FAK) is known to regulate cell adhesion, migration, and mechanotransduction in focal adhesions (FAs). However, studies on how FAK activity is regulated in the plasma membrane microdomains according to the composition of extracellular matrix (ECM) proteins are still lacking. A genetically encoded fluorescence resonance energy transfer (FRET)-based biosensor can provide useful information on the activity of intracellular signals with high spatiotemporal resolution. In this study, we analyzed the FAK activities in lipid raft (detergent-resistant membrane) and non-lipid raft (non-detergent-resistant membrane) microdomains using FRET-based membrane targeting FAK biosensors (FAK-Lyn and FAK-KRas biosensors) under four different ECM protein compositions: glass, type 1 collagen, fibronectin, and laminin. Interestingly, FAK activity in response to laminin in a lipid raft microdomain was lower than that in other ECM conditions. Cells subjected to fibronectin showed higher FAK activity in a lipid raft microdomain than that in a non-lipid raft microdomain. Therefore, this study demonstrates that the FAK activity can be distinctively regulated according to the ECM type and the environment of the plasma membrane microdomains.
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KEYWORD
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Biosensor, extracellular matrix, focal adhesion kinase, FRET, single cell imaging
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