Evaluation of the Number of Agonist Molecules Needed to Activate a Ligand-Gated Channel from the Current Rising Phase

Abstract

We propose a new method for calculating the number of agonist binding sites ( n) in ligand-gated receptor channels from the initial phase of the current. This method is based on the fact that the relation between the current ( I) and its first-time derivative ( I′) at the beginning of the current reflects the number of transitions that lead to channel opening. We show that, for constant agonist concentration, the above relationship at t → 0 provides the number of steps leading to channel opening. When the agonist concentration is not constant but rather increases linearly with time, the corresponding value can be obtained using a slightly modified procedure. The analytical results were compared with computer simulations and a good match between the two was obtained. The theoretical procedure was then validated experimentally using the nicotinic receptor, because, for this receptor, the number of binding sites is well established. Indeed, the expected number of two binding sites was obtained. The method was then tested for the quisqualate-type glutamate receptor channel from the opener muscle of crayfish. The number of this receptor's binding sites is not fully resolved. Our results suggest that, for this glutamate receptor as well, two binding sites must be occupied to open the channel.