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. 2018 Aug 27;115(37):E8642–E8651. doi: 10.1073/pnas.1717594115

Fig. 7.

Fig. 7.

Actin filaments increase the FRET efficiency between dyes on both Arp2/Arp3 and ArpC1/ArpC3. Ensemble ETeff measurements of 20 nM Arp2/3 complex labeled (blue) with FlAsH on ArpC3 and Alexa 568 on ArpC1 or (orange) with FlAsH on Arp2 and Alexa 568 on Arp3. The measurements are differences between the ETeff values of Arp2/3 complex alone and with the each ligand. Conditions: 50 mM KCl, 1 mM MgCl2, 1 mM EGTA, 0.1 mM ATP, 1 mM DTT, and 10 mM imidazole, pH 7.0, unless indicated otherwise. (A) Titration of Arp2/3 complex with Wsp1p-VCA. (B) Titration of Arp2/3 complex with polymerized actin with overnight incubation. (C) Titration of Arp2/3 complex and 10 μM polymerized actin with Wsp1p-VCA. (D) Interleaved scatter plot of differences in mean ETeff values (±1 SD) of labeled Arp2/3 complex alone and in the presence of 2 mM ATP and with 2 mM ATP and various ligands: 20 µM Wsp1p-CA, 20 µM Wsp1p-VCA, 20 µM GST-VCA, 2 µM actin-VCA (actin monomer covalently linked to VCA), 10 µM polymerized actin with 10 µM VCA, or 20 µM polymerized actin. Numbers of repetitions are in parentheses.