The three-finger toxin MTalpha is a selective alpha(2B)-adrenoceptor antagonist.

Muscarinic toxins (MTs) are three-finger folded peptides isolated from mamba snake venoms. In this report we describe a selective antagonistic interaction of MTalpha with the human alpha(2B)-adrenoceptor. In a functional assay, measuring the alpha(2B)-adrenoceptor-induced increase in intracellular [Ca(2+)], we found that both venomous MTalpha and synthetic MTalpha inhibited the response in a concentration-dependent way. MTalpha did not affect the responses of alpha(2A)-, alpha(2C)-, alpha(1A)- or alpha(1B)-adrenoceptors. To further explore the binding of MTalpha to the alpha(2B)-adrenoceptor, we performed ligand binding experiments on Sf9 cell homogenates with [(3)H]RX821002 as reporter ligand. MTalpha bound to the receptor rather slowly requiring about 60 min to reach equilibrium. In equilibrium binding experiments, MTalpha displaced the radioligand with an IC(50) of 3.2 nM, but was not able to displace all bound radioligand. Using a saturation binding protocol, we found that MTalpha suppressed the maximum binding without any greater impact on the affinity of the radioligand, indicating a non-competitive mode of inhibition. The toxin bound reversibly to alpha(2B)-adrenoceptor, but extensive washing was needed for full recovery of binding sites at high toxin concentrations. Surprisingly, MTalpha did not affect [(3)H]-N-methylscopolamine binding to the muscarinic receptor subtypes at concentrations found to fully block alpha(2B)-adrenoceptors, showing that the toxin is a more potent antagonist for the alpha(2B)-adrenoceptor than for muscarinic receptors. These findings should open up new views in terms of selective adrenoceptor drug design as well as in elucidation of alpha(2)-adrenoceptor physiology. Copyright 2010 Elsevier Ltd. All rights reserved.