哺乳類細胞における調節可能なアクチン-ミオシン収縮性と近接性ビオチン化を用いた力学的感受性核タンパク質の同定プロトコル

Mechanical forces influence a range of cellular behaviors; however, how these forces are sensed and converted into biochemical changes remains incompletely understood. A key aspect of mechanotransduction is the regulation of subcellular protein localization. Here, we present a protocol describing the engineering of cell lines with tunable actomyosin contractility combined with a proximity biotinylation strategy confined to the nucleus followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. This approach allows the identification of proteins whose nuclear localization is controlled by changes of actomyosin contractility. For complete details on the use and execution of this protocol, please refer to Tseng et al.¹.