Figure 2.
Stereochemical and charge effects on the interaction of of the LFN N-terminus with PA pore. a) Chemical framework of SSv1-SSv4. b) The fraction ion conductance block of PA pore by LFN variants at Δ Ψ = 20 mV (Δ Ψ = Ψcis − Ψtrans, where Ψtrans ≡ 0). For the procedure used to determine the fraction ion conductance block shown in Figure 2 and Figure 3 see the supplementary information. Each blocking experiment was repeated three times. c) Acid triggered translocation of LFN variants through PA pore in response to a pH gradient of ~2 units (cis pH 5.5; trans pH 7.5) at Δ Ψ = 20 mV. d) Translocation of LFN-DTA variants through PA pore in response to a pH gradient of ~2 units (cis pH 5.5; trans pH 7.5) at Δ Ψ = 20 mV. e) Fraction protein synthesis inhibition by LFN-DTA variants in CHO-K1 cells. LFN-DTA variants were incubated at the indicated concentration for 15–17 minutes at 37°C and 5 % CO2. Then, 3H-leucine was added, and after 1 hour the amount of tritium incorporated into cellular protein was measured. The data shown are the average of three experiments. Full experimental details are described in the supplementary information.