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Fig. 5. Xin repeats bind directly to actin filaments. Bacterially expressed XR1-16 (A-C) and its truncated variants XR1-6 (D-F), XR1-3 (G,H) and XR3-4 (I) were purified and their abilities to bind skeletal-muscle F-actin were investigated using high-speed co-sedimentation assays. The positions of actin and recombinant truncated Xin are marked with an asterisk and an arrow, respectively. The numbers on top of each lane indicate the amount of recombinant protein that was added to 5 µM actin. In H, the pellet was dissolved in less sample buffer (25%) to be able to visualize precipitated XR1-3 clearly. XR1-16, XR1-6 and XR1-3 are precipitated together with F-actin, whereas the two-repeat construct XR3-4 remains in the supernatant. Binding of the full Xin-repeat region to 1 µM F-actin is saturated with approximately 0.25 µM XR1-16 (C), whereas complete saturation of binding of XR1-6 to 1 µM F-actin is reached with approximately 0.5 µM recombinant protein (F).

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