Pav binds to actin through its head region. (A) Schematic diagrams illustrating the protein domains of Pav and different pieces of Pav used in bundling and cosedimentation assays. (B–E”) Stabilized actin and/or MTs were bundled and cross-linked by Pav-head (B–B”), Pav-head + stalk (C–C”), Pav-stalk + tail (D and D’), and Pav-tail (E and E’). (F) High-speed cosedimentation assay of actin in the presence of different Pav protein pieces. The percentage of Pav protein segregating with the pellets or remaining in the supernatant is shown at the bottom of gel. (G) Quantification of Pav proteins pelleting in the presence or absence of actin. (H–H”) Stabilized actin and/or MTs were incubated with Pav^DEAD. (I–L”) Stabilized actin or MTs were incubated with CapuFH2 to bundle actin or MTs, followed by the addition of GFP-Pav (I–J”) or GFP-Pav^DEAD (K–L”). Final protein concentrations for bundling assays: Pav-head, 1 µM; Pav-stalk + tail, 1 µM; Pav-head + stalk, 300 nM; Pav-tail, 2 µM; Pav-Pav^DEAD, 200 nM; CapuFH2, 500 nM; GFP-Pav, 3 nM; and GFP-Pav^DEAD, 3 nM. Final protein concentrations for cosedimentation assays: Pav-head, 300 nM; Pav-stalk + tail, 300 nM; Pav-head + stalk, 200 nM; Pav-tail, 800 nM; Pav^DEAD-head, 300 nM; BSA, 2 µM; and actin, 2 µM. Scale bar: 30 µm.