Relationship between imidazoline/guanidinium receptive sites and monoamine oxidase A and B

Imidazoline binding sites or imidazoline/guanidinium receptive sites (IGRS) recognize bioactive endogenous substances and a variety of pharmacologically active compounds containing imidazoline or guanidinium moieties. The family of imidazoline binding proteins consists of multiple membrane-associated proteins that differ in their tissue/subcellular localization, M(r) and ligand recognition properties. Two of the imidazoline binding proteins are identical to the mitochondrial enzyme monoamine oxidase (MAO) A and B isoforms, which contain imidazoline binding domains distinct from the enzyme active site. The relationship between the imidazoline binding proteins and monoamine oxidases was further characterized in the present report using a covalent probe (2-[3-azido-4[125I]iodophenoxy] methyl imidazoline, [125I]-AZIPI) to label the imidazoline binding proteins in different species and following transient expression of MAO- A and -B in COS 7 cells. Species homologues of MAO-A and -B in rat and human differ in their apparent molecular weight by approximately 2000 Da. In rat and human liver [125I]-AZIPI identified peptides with apparent molecular weights similar to those of the species homologues of MAO. Peptides of M(r) approximately 63,000 (MAO-A) and approximately 59,000 (MAO-B) were also photolabeled in membranes prepared from COS-7 cells transfected with human cDNA clones encoding MAO-A or -B. Additional experiments indicate that the imidazoline binding domains on MAO-A and -B exhibit different ligand recognition properties. The covalent labeling of human liver MAO-B was more sensitive than that of placenta MAO-A to inhibition by the imidazoline 2-(4,5-dihydroimidaz-2-yl)-quinoline (BU224). These data indicate that the A and B isoforms of MAO possess imidazoline binding domains that differ in their ligand recognition properties. Allosteric regulation of the activity of MAO via the imidazoline binding domains may be of significance in various disease states associated with elevated enzyme expression or in which the enzyme is a therapeutic target.