Acetylcholine receptor from Narke japonica electroplax exhibits a fluorescence change upon binding of snake neurotoxins. This fluorescence change primarily arises from the conformational change of the acetylcholine receptor and reflects the binding process of the toxin with the receptor. The time dependence of the fluorescence change has been monitored for 28 short neurotoxins and 8 long neurotoxins by using a stopped-flow technique. The steady-state fluorescence change is of the same order of magnitude for the short neurotoxins but varies among the long neurotoxins. Nha 10, a short neurotoxin with weak neurotoxicity, causes no fluorescence change in the receptor but can still bind to the receptor with sufficiently high affinity. The substitution of the conserved residue Asp-31 to Gly-31 in Nha is probably responsible for the reduced neurotoxicity. The rate constants for the binding of the neurotoxins to the receptor have been obtained by analyzing the transient fluorescence change. The rate constants show surprisingly a wide range of distribution: (1.0-20.5) X 10(6) M-1 s-1 for short neurotoxins and (0.26-1.9) X 10(6) M-1 s-1 for long neurotoxins. Examination of the relationship between the rate constants of fluorescence change of the short neurotoxins and their amino acid sequences, thermal stability, hydrogen-deuterium exchange behavior, overall net charge, etc. reveals the following. Positive charges on the side chains of residues 27 and 30 and overall net charge of the neurotoxin govern the magnitude of the binding rate of the neurotoxin with the receptor.