Charybdotoxin unbinding from the mKv1.3 potassium channel: A Combined computational and experimental study
Rudiger Ettrich1,2
1Institute of Nanobiology and Structural Biology of GCRC, Academy of Sciences of the Czech Republic, Zamek 136, CZ-37333 Nove Hrady, Czech Republic
2Faculty of Sciences, University of South Bohemia, Zamek 136, CZ-37333 Nove Hrady, Czech Republic
Charybdotoxin, belonging to the group of so-called scorpion toxins, is a short
peptide able to block many voltage-gated potassium channels, such as mKv1.3, with high affinity. We use a
reliable homology model based on the high-resolution crystal structure of the
94% sequence identical homolog Kv1.2 for charybdotoxin
docking followed by molecular dynamics simulations to investigate the mechanism
and energetics of unbinding, tracing the behavior of
the channel protein and charybdotoxin during
umbrella-sampling simulations as charybdotoxin is
moved away from the binding site [1, 2]. The potential of mean force is
constructed from the umbrella sampling simulations and combined with Kd and free energy values gained experimentally using the patch-clamp
technique to study the free energy of binding at different ion concentrations
and the mechanism of the charybdotoxin-mKv1.3 binding process. A possible
charybdotoxin binding mechanism is deduced that includes an initial hydrophobic
contact followed by stepwise electrostatic interactions and finally
optimization of hydrogen-bonds and salt-bridges. T
R.E. acknowledges the contributions of Morteza Khabiri, Azadeeh Nikouee and Stephan Grissmer and support from the Czech Science Foundation, Grant 13-21053S.
[1] Khabiri, M.; Nikouee, A.; Cwiklik, L.; Grissmer, S.; Ettrich, R.
Journal of Physical Chemistry B 2011 115 (39): 11490–11500.[2] Nikouee, A.; Khabiri, M.; Grissmer, S.; Ettrich, R. Journal of Physical Chemistry B 2012, 116 (17): 5132-5140.