Panel A shows two gel images and a bar graph. The gels display the presence of Mical, Actin, and Espin in supernatant (S) and pellet (P) fractions. The bar graph quantifies the percentage of pelleted actin, showing a significant reduction when Mical and NADPH are added. Panel B presents two gel images and a bar graph. The gels illustrate the presence of Mical, Fascin, Actin, and Espin in supernatant and pellet fractions. The bar graph indicates a substantial decrease in pelleted actin with the addition of Mical and NADPH. Panel C includes two gel images and a bar graph. The gels show the distribution of Fascin, Actin, and Espin in supernatant and pellet fractions under different conditions. The bar graph quantifies the percentage of pelleted actin, demonstrating a decrease when Mical and NADPH are present.
Mical robustly disassembles complex different protein-bundled F-actin structures. (A) Mical/NADPH disassembles espin-bundled F-actin. In low-speed pelleting assays, a majority of espin-bundled F-actin is present in the pellets (left). Addition of Mical and NADPH robustly reduces the amount of F-actin in the pellets (blue arrowhead, graph). [F-actin] = 2.5 μM; [Espin] = 1.25 μM; [Mical] = 0.6 μM; [NADPH] = 100 μM. Mean values ± SEM. n = 2 independent experiments/conditions. (B) Mical/NADPH disassembles fascin- and espin-bundled F-actin. In low-speed pelleting assays, a majority of fascin + espin-bundled F-actin is present in the pellets (left), but the addition of Mical and NADPH decreases strongly the pelleted F-actin (blue arrowhead, graph). [F-actin] = 2.5 μM; [Fascin] = 1.25 μM; [Espin] = 1.25 μM; [Mical] = 0.6 μM; [NADPH] = 100 μM. Mean values ± SEM. n = 2 independent experiments/conditions. (C) In high-speed pelleting assays, in all cases a majority of actin is present in the pellets (F-actin), but the addition of Mical and NADPH decreases the pelleted F-actin (plot on the right) and disassembles it down to G-actin (gel on the left). [F-actin] = 2.5 μM; [Fascin] = 1.0 μM for fascin only bundles; [Espin] = 1.0 μM for espin only bundles; [Fascin] = 0.5 μM and [Espin] = 0.5 μM for fascin–espin co-bundles; [Mical] = 0.6 μM; [NADPH] = 100 μM. Mean values ± SEM. n = 3 independent experiments/conditions. Source data are available for this figure: SourceData F3.