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Proteins, Expression, Isolation and Analysis> | The cell lysate was centrifuged at ~50,000 g for 30 min, and the resulting supernatant was then loaded onto anti-Flag beads (GenScript) followed by a 1-h incubation. After extensive washing, the target protein was eluted with the lysis buffer without Triton X-100, supplemented with Flag peptide (500 μg/ml). The prepared samples were separated by 4-12% SDS-PAGE (GenScript, M41212C) and transferred to a polyvinylidene fluoride membrane (Millipore, IPVH00010). | Get A Quote |
Microtubule organization in cells relies on targeting mechanisms. Cytoplasmic linker proteins (CLIPs) and CLIP-associated proteins (CLASPs) are key regulators of microtubule organization, yet the underlying mechanisms remain elusive. Here, we reveal that the C-terminal domain of CLASP2 interacts with a common motif found in several CLASP-binding proteins. This interaction drives the dynamic localization of CLASP2 to distinct cellular compartments, where CLASP2 accumulates in protein condensates at the cell cortex or the microtubule plus end. These condensates physically contact each other via CLASP2-mediated competitive binding, determining cortical microtubule targeting. The phosphorylation of CLASP2 modulates... More