Extended Data Fig. 2: Activation of AC5 by Gβγ and purification of an AC5–Gβγ complex.
From: Structure of adenylyl cyclase 5 in complex with Gβγ offers insights into ADCY5-related dyskinesia
(A) AC activity of purified AC5 was determined in the presence of 10 mM MgCl2 and 0.5 mM ATP with or without 250 nM Gαs·GTPγS, in the absence or presence of 400 nM Gβ1γ2. Data presented are the mean ± SD. Statistical significance was assessed using an unpaired Welch’s t-test (two-sided) across at least five independent experiments. (B) The AC5–Gβγ complex was purified using anti-FLAG M2 affinity resin and injected onto a Superose 6 Increase 3.2/300 column equilibrated with 50 mM HEPES pH 8, 100 mM NaCl, 10 mM MgCl2, and 0.01 % LMNG. Peak fractions highlighted in pink were pooled, concentrated, and (C) analyzed by SDS-PAGE (note that Gγ2 runs off the gel).
