Figure 1. Interactions between Kap114,... | Rockefeller University Press

Figure 1.

$Interactions between Kap114, Nap1, H2A-H2B, and RanGTP. (A) Organization schematics of the Nap1 (cyan), H2A (yellow), H2B (magenta), and Kap114 (gray, long loops labeled) polypeptides. (B) Pull-down assay with immobilized MBP-Kap114 (1 µM) ± equimolar Nap12 ± RanGTP. (C) SEC-MALS analysis of equimolar Kap114, Nap12, and H2A-H2B mixtures without (orange) and with RanGTP (green). Left panel: differential refractive index (dRI, left y-axis, thin lines) and molecular mass (kDa, right y-axis, thick lines) traces, with theoretical masses of the Kap114•H2A-H2B (K•H), Kap114/Nap12/H2A-H2B (K/N2/H), and Kap114/Nap12/H2A-H2B/RanGTP (K/N2/H/R) complexes marked with dashed lines. Right panels: Coomassie-stained SDS-PAGE of peak fractions. When RanGTP is absent, the presence of a minor peak that matches K•H suggests the likely presence of some free Nap1 oligomers. Controls in Figs. S1 and S2. Source data are available for this figure: SourceData F1.$

Interactions between Kap114, Nap1, H2A-H2B, and Ran ^GTP . (A) Organization schematics of the Nap1 (cyan), H2A (yellow), H2B (magenta), and Kap114 (gray, long loops labeled) polypeptides. (B) Pull-down assay with immobilized MBP-Kap114 (1 µM) ± equimolar Nap1₂ ± Ran^GTP. (C) SEC-MALS analysis of equimolar Kap114, Nap1₂, and H2A-H2B mixtures without (orange) and with Ran^GTP (green). Left panel: differential refractive index (dRI, left y-axis, thin lines) and molecular mass (kDa, right y-axis, thick lines) traces, with theoretical masses of the Kap114•H2A-H2B (K•H), Kap114/Nap1₂/H2A-H2B (K/N₂/H), and Kap114/Nap1₂/H2A-H2B/Ran^GTP (K/N₂/H/R) complexes marked with dashed lines. Right panels: Coomassie-stained SDS-PAGE of peak fractions. When Ran^GTP is absent, the presence of a minor peak that matches K•H suggests the likely presence of some free Nap1 oligomers. Controls in Figs. S1 and S2. Source data are available for this figure: SourceData F1.

This Feature Is Available To Subscribers Only