Extended Data Fig. 6: CO2 blocks BODIPY FL ATP-γ-S binding to PII from Synechocystis sp., related to Fig. 5.
From: Chemoproteomic identification of CO2-dependent lysine carboxylation in proteins
a, FP equilibrium dissociation binding of BODIPY FL ATP-γ-S to WT and K90A SsPII. b-c, FP equilibrium competition binding experiments for WT (b) and K90A SsPII (c) in the presence of varying concentrations of small molecules (2-OG, GTP, UDP, ADP, ATP). d, Time-dependant effect of KOCN on FP equilibrium probe binding to WT and K90A SsPII. e, FP equilibrium binding of BODIPY FL ATP-γ-S following pre-incubation of SsPII in the presence and absence of KOCN followed by jump dilution and desalting to remove residual KOCN. In (a-e), all data is presented as mean values ± SD from n = 2 independent experiments. f, FP equilibrium competition binding experiments for WT and K90A SsPII in the presence of varying concentrations of HCO3−. Data in (f) are presented as mean values ± SD from n = 4 independent experiments. In (b) and (f), KD’s for competitor compounds were calculated from IC50 values using a standard FP equilibrium binding formula as previously described59. KD(CO2) was determined from KD using standard equations for carbonate equilibria as previously described60.
