Figure 4.
ATL2 C-terminus impairs G domain–mediated tethering and GTP hydrolysis, but not GTP binding.(A) Mant-GTP binding by FL ATL2, ATL2(1–565), ATL2(1–547), and K107A ATL2(1–547). Baseline fluorescence was established for 40 s followed by addition of ATL2 with the first reading after 20 s, an average of ≥5 individual traces (±SEM). (B) Tethering monitored as the change in 405-nm absorbance after the addition of 2 mM GTP. (C) Reversibility of GTP-dependent tethering by ATL2(1–547). 120 s after the initial addition of buffer or GTP as in B, EDTA was added to 18 mM. (D) Steady-state GTPase activity. Average (±SEM) of at least three independent measurements. All assays used ATL2 proteins incorporated at a 1:1,000 protein/lipid ratio.

ATL2 C-terminus impairs G domain–mediated tethering and GTP hydrolysis, but not GTP binding. (A) Mant-GTP binding by FL ATL2, ATL2(1–565), ATL2(1–547), and K107A ATL2(1–547). Baseline fluorescence was established for 40 s followed by addition of ATL2 with the first reading after 20 s, an average of ≥5 individual traces (±SEM). (B) Tethering monitored as the change in 405-nm absorbance after the addition of 2 mM GTP. (C) Reversibility of GTP-dependent tethering by ATL2(1–547). 120 s after the initial addition of buffer or GTP as in B, EDTA was added to 18 mM. (D) Steady-state GTPase activity. Average (±SEM) of at least three independent measurements. All assays used ATL2 proteins incorporated at a 1:1,000 protein/lipid ratio.

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