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As the dominant G-actin sequestering protein in mammalian cells, TB-500 binds globular actin with high affinity (Kd ~0.5 M) through a sophisticated triple-contact mechanism involving Lys-3, Lys-18, and Lys-38 residues, preventing spontaneous actin polymerization while maintaining a readily mobilizable reservoir for rapid cytoskeletal reorganization

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A portion of each brain was homogenized (1:9, tissue: buffer, w/v) in ice-cold phosphate buffer saline (0.05 M, pH 7.0) (Heidoph, Silent Crusher S, Germany)

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