Figure 5. The C-terminal region of Bnr1 is not required for Bud6 binding. (A) Quantitative GST pull-down assays using immobilized GST-Bud6(L) or GST-Bud6(S) and soluble C-Bni1Δtail (left) or C-Bnr1Δtail (right). (B) 2 µM monomeric actin was assembled alone, with 2 nM C-Bnr1Δtail, or with 2 nM C-Bnr1Δtail + 200 nM Bud6(L). Reactions were performed in the absence (left) or presence (right) of 5 µM yeast profilin. (C) Quantitative GST pull-down assays as in A using immobilized GST-Bud6(S) and soluble Bnr1 FH2. For each panel of this figure, the data shown are single representative examples from two independent experiments. The lines in A and C are fits to the equation B = a × c/(c + K), in which B is the fraction of C-Bnr1Δtail (A) or Bnr1 FH2 (C) bound, a is the amplitude of the binding interaction, c is the molar concentration of GST-Bud6, and K is the binding affinity (i.e., Kd). AU, arbitrary unit.
Figure 5.

The C-terminal region of Bnr1 is not required for Bud6 binding. (A) Quantitative GST pull-down assays using immobilized GST-Bud6(L) or GST-Bud6(S) and soluble C-Bni1Δtail (left) or C-Bnr1Δtail (right). (B) 2 µM monomeric actin was assembled alone, with 2 nM C-Bnr1Δtail, or with 2 nM C-Bnr1Δtail + 200 nM Bud6(L). Reactions were performed in the absence (left) or presence (right) of 5 µM yeast profilin. (C) Quantitative GST pull-down assays as in A using immobilized GST-Bud6(S) and soluble Bnr1 FH2. For each panel of this figure, the data shown are single representative examples from two independent experiments. The lines in A and C are fits to the equation B = a × c/(c + K), in which B is the fraction of C-Bnr1Δtail (A) or Bnr1 FH2 (C) bound, a is the amplitude of the binding interaction, c is the molar concentration of GST-Bud6, and K is the binding affinity (i.e., Kd). AU, arbitrary unit.

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