THE USE OF IRREVERSIBLE LIGANDS TO INACTIVATE RECEPTOR SUBTYPES - 4-DAMP MUSTARD AND MUSCARINIC RECEPTORS IN SMOOTH-MUSCLE

Irreversible ligands are useful tools for investigating the function o f receptor subtypes in various physiological processes. The mechanism for alkylation involves the formation of a reversible receptor complex followed by a covalent reaction. The extent of receptor alkylation is determined by the dissociation constant of the reversible complex and the rate constant for conversion to the covalent complex. Selectivity can be achieved if the irreversible ligand exhibits a difference in i ts dissociation constants for receptor subtypes. Selective alkylation can also be achieved using a selective competitive inhibitor to protec t the desired receptor subtype. By using the non-M-2-selective irrever sible antagonist, 4-DAMP mustard, in combination with the competitive M-2-selective antagonist, AF-DX 116, it has been possible to achieve a highly selective inactivation of all non-M-2 subtypes of the muscarin ic receptors in smooth muscle and has enabled the discovery of the fun ctional role of M-2 receptors in smooth muscle.