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. Author manuscript; available in PMC: 2015 Jun 6.
Published in final edited form as: J Am Chem Soc. 2015 Apr 23;137(17):5793–5797. doi: 10.1021/jacs.5b01520

Figure 2.

Figure 2

Binding of ligands to AlkB-Fe++ confined inside ClyA-AS. a, Typical ligand-induced blockades to individual AlkB-Fe++ enzymes confined inside ClyA-AS at −60 mV. The ligand used is shown on the right of the trace. The bound Level 1 current levels (L1O, L1N, L1S) are represented by the blue dashed lines. The substrate concentration was 0.6 mM for 2-OG, 0.6 mM for N-OG and 2 mM for SUC binding to wild type AlkB-Fe++, and 7.2 mM for 2-OG binding to N120D-AlkB-Fe++. b, Left: Dissociation rate constants (koff) as a function of the ligand concentration at −60 mV. Right: Event frequency (1/τon) as a function of the ligand concentration at −60 mV. 2-OG is shown in yellow squares, SUC in black circles and N-OG in red triangles. All current traces were collected by applying a Bessel-low pass filter with a 2 kHz cut-off and sampled at 10 kHz. An additional Bessel 8-pole filter with 50 Hz cut-off was digitally applied to the current traces. All recordings were carried out in 150 mM NaCl, 15 mM Tris HCl pH 8.0, at 28°C, and the ligands were added to the cis compartment. Errors are given as standard deviations.