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KMID : 0613820230330020191
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Journal of Life Science
2023 Volume.33 No. 2 p.191 ~ p.198
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Comparative Analysis of Src Activity in Plasma Membrane Subdomains via Genetically Encoded FRET Biosensors
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Choi Gyu-Ho
Jang Yoon-Kwan
Suh Jung-Soo
Kim Heon-Su
Ahn Sang-Hyun
Kim Tae-Jin
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Abstract
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As a member of the focal adhesion complex of the plasma membrane, Src is a nonreceptor tyrosine kinase that controls cell adhesion and motility. However, how Src activity is regulated in the plasma membrane microdomain in response to components of the extracellular matrix (ECM) remains unclear. This study compared and investigated the activity of Src in response to three representative ECM proteins: collagen type 1, fibronectin, and laminin. Genetically encoded FRET-based Src biosensors for plasma membrane subdomains were used. FRET-based biosensors allow the real-time analysis of protein activity in living cells based on their high spatiotemporal resolution. The results showed that Src activity was maintained at a high level under all ECM conditions of the lipid raft, and there was no significant difference between the ECM conditions. In contrast, Src activity was maintained at a low level in the non-lipid raft membrane. In addition, the Src activity of lipid rafts remained significantly higher than that of non-lipid raft regions under the same ECM conditions. In conclusion, this study demonstrates that Src activity can be controlled differently by lipid rafts and non-lipid raft microdomains.
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KEYWORD
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Biosensor, extracellular matrix, FRET, single-cell imaging, src
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