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. 2021 Jan 13;10:e64624. doi: 10.7554/eLife.64624

Figure 4. BART product inhibition is relieved by ARL3 effectors which mediate its turnover.

Figure 4.

(A) Nucleotide exchange of ARL3 was assayed in a multiple turnover experiment with a limiting addition of BART. Fluorescence polarisation was measured following nucleotide addition for a mixture of 20 μM ARL3mantGDP, 20 μM ARL13B, and 1 μM BART in the presence of either 150 μM GppNHp (red) or 150 μM GDP (blue). (B) Fluorescence polarisation was measured for a mixture of 5 μM ARL3mantGDP, 100 μM GppNHp, and 1 μM ARL13B18-278 (grey). The measurements were repeated in the presence of 5 μM UNC119A (yellow), 2 μM BART (blue), or a mixture of UNC119A and BART at the aforementioned concentrations (orange). All reactions were initiated by the addition of GppNHp. (C) SPR sensorgrams of ARL3GppNHp (640 nM) bound to immobilised GST-BART undergoing nine washing steps (black arrows) with either running buffer (red), or with 2 μM PDEδ (blue). The data was fitted to a single exponential equation using GraFit, and the fitted curves are shown on the right, along with the corresponding rate constants.

Figure 4—source data 1. Source data for Figure 4a.
Figure 4—source data 2. Source data for Figure 4b.
Figure 4—source data 3. Source data for Figure 4c.
elife-64624-fig4-data3.xlsx (975.1KB, xlsx)