Figure 2.
Strong interaction between Kip2 and Bik1 requires the C-terminal tail of Kip2. (A) Schematic representations of protein constructs used in this study. Zn, Zinc knuckle domain. (B) SEC-MALS experiments of Kip2-NMD (green; calculated molecular mass of the monomer: 23 kD), tubulin (dark gray; calculated molecular mass of αβ-tubulin: 110 kD), and a mixture of Kip2-NMD and tubulin (blue). Note that the molecular mass distribution under the Kip2-NMD/tubulin mixture elution profile is in between the ones of the Kip2-NMD dimer and the tubulin heterodimer, indicating no interaction between the two proteins. (C) Coomassie-Blue-stained SDS-PAGE of microtubule pelleting assays using Taxol-stabilized microtubules mixed without and with Kip2-NMD. S, supernatant; P, pellet. The supernatant lane of the Kip2-NMD alone sample is also shown in Fig. S1 E. (D) SEC-MALS analyses of Bik1-CC (green), Kip2-NMD (dark blue), and a mixture of Bik1-CC and Kip2-NMD (purple). (E) SEC-MALS analyses of Bik1-CC (green), Kip2-NMD-ΔT (light blue), and a mixture of Bik1-CC and Kip2-NMD-ΔT complex (orange). The UV absorption at 280 nm and the molecular masses across the peak determined by MALS are plotted. Source data are available for this figure: SourceData F2.

Strong interaction between Kip2 and Bik1 requires the C-terminal tail of Kip2. (A) Schematic representations of protein constructs used in this study. Zn, Zinc knuckle domain. (B) SEC-MALS experiments of Kip2-NMD (green; calculated molecular mass of the monomer: 23 kD), tubulin (dark gray; calculated molecular mass of αβ-tubulin: 110 kD), and a mixture of Kip2-NMD and tubulin (blue). Note that the molecular mass distribution under the Kip2-NMD/tubulin mixture elution profile is in between the ones of the Kip2-NMD dimer and the tubulin heterodimer, indicating no interaction between the two proteins. (C) Coomassie-Blue-stained SDS-PAGE of microtubule pelleting assays using Taxol-stabilized microtubules mixed without and with Kip2-NMD. S, supernatant; P, pellet. The supernatant lane of the Kip2-NMD alone sample is also shown in Fig. S1 E. (D) SEC-MALS analyses of Bik1-CC (green), Kip2-NMD (dark blue), and a mixture of Bik1-CC and Kip2-NMD (purple). (E) SEC-MALS analyses of Bik1-CC (green), Kip2-NMD-ΔT (light blue), and a mixture of Bik1-CC and Kip2-NMD-ΔT complex (orange). The UV absorption at 280 nm and the molecular masses across the peak determined by MALS are plotted. Source data are available for this figure: SourceData F2.

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