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. 2022 Jan 14;11:e72202. doi: 10.7554/eLife.72202

Figure 1. Cytoskeleton-Associated Protein 2 (CKAP2) is an intrinsically disordered protein that increases microtubule formation.

(a) Schematic of mmCKAP2 protein domains and disorder prediction (Dosztányi et al., 2005; Obradovic et al., 2003). (b) Circular dichroism of 4.5 μM CKAP2. (c) Coomassie Blue-stained sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) gel of 1 μg of purified recombinant CKAP2 constructs and 5 μg of purified tubulin. (d) Light scattering assay (turbidity) schematic and data following microtubule formation as apparent absorbance (Abs) over time for 8 µM tubulin with increasing concentrations of CKAP2-mNeonGreen (CKAP2-mNG) (n = 1). (e) Turbidity data for tubulin alone (blue) and addition of CKAP2-mNG (n = 3, mean ± standard deviation [SD]).

Figure 1—source data 1. Protein purification gel.

Figure 1.

Figure 1—figure supplement 1. Cytoskeleton-Associated Protein 2 (CKAP2) is an intrinsically disordered protein that increases microtubule assembly.

Figure 1—figure supplement 1.

(a) Comparison of amino acid sequence conservation for CKAP2, TPX2, and chTOG between human (hs, Homo sapiens), mouse (ms, Mus musculus), and African clawed frog (xl, Xenopus laevis) and amino acid sequence alignment for CKAP2. Dark grey shading represents sequence identities, light grey shading sequence similarities. KEN box (green) and CKAP2_C domain (magenta) are underlined. (b) Turbidity assay with CKAP2-mNeonGreen (mNG) no-His construct (n = 1). (c) Turbidity assay with CKAP2 unlabelled (no-mNG) (n = 1). (d) Turbidity assay performed with CKAP2-mNG at 4°C (n = 2).