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. Author manuscript; available in PMC: 2022 Jun 2.
Published in final edited form as: Nat Chem Biol. 2021 Dec 2;18(4):360–367. doi: 10.1038/s41589-021-00900-9

Extended Data Fig. 3: Analysis of apoptolidin A binding mode via photoaffinity labeling, and enzymatic assays.

Extended Data Fig. 3:

a, Concentration response-curves showing inhibition of ATPase activity in purified yeast FOF1 ATP synthase, measured using PK/LDH coupled assay; b, Gel-based analysis of Apop A competition against Apop A-PA at a fixed concentration; c, Gel-based analysis of Apop A-PA in the presence of known ATP synthase inhibitors Aurovertin B (Auro B), Efrapeptin F (Efra F), Citreoviridin (Citr), and Oligomycin A (Oligm); d, Gel-based profiling of Apop A-PA in MV-4–11 cells treated with or without uncoupling agents (1 μM). e, Analysis of ADP concentration dependence on inhibition of ATP synthesis by Apop A in isolated mouse liver mitochondria using HK/G6PDH coupled assay; f, Analysis of ATP concentration dependence on inhibition of ATP hydrolysis by Apop A and Ammo A in isolated yeast F1 ATPase using a PK/LDH coupled assay; g, confirmation of ATPIF1 KO in two independent K562 clones after knockout using PX458 with two independent ATP5IF1 targeting gRNAs; h, Gel-based profiling of membrane potential dependent adduction of Apop A-PA in WT or IF1 KO K562 cells; i, comparison of ammocidin bound (left) to IF1 bound F1 ATPase.