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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

In vitro and in vivo effect of BU99006 (5-isothiocyanato-2-benzofuranyl-2-imidazoline) on I(2) binding in relation to MAO: Evidence for two distinct I(2) binding sites.

BU99006 is an irreversible I(2) ligand which selectively inactivates I(2) binding sites, making it an ideal tool with which to study I(2) site mechanism. We sought to determine the effects of BU99006 on I(2) binding in relation to monoamine oxidase (MAO), and the time course of these effects. In vitro, rat brain membranes that were pre-treated with 10muM BU99006 showed no change in MAO activity, despite suffering a significant reduction in [(3)H]2BFI binding (52.5+/-19.6 to 8.5+/-3.8fmol mg(-1), 84%). Furthermore, reversible I(2) ligands 2BFI and BU224 were able to inhibit MAO, whether treated with BU99006 or not. In vivo, a 5mg kg(-1) i.v. dose of BU99006 in rats rapidly reduced [(3)H]2BFI binding with similar magnitude (85%, maximal reduction after 20min), without effect on either MAO activity or the specific binding of selective MAO-A and MAO-B radioligands. Moreover, following this irreversible treatment, recovery of central [(3)H]2BFI binding occurred with a rapid half-life of 4.3h in rat brain (2.0h in mouse), which is not consistent with a site on MAO. These data indicate that the high affinity site which is occupied by [(3)H]2BFI and irreversibly binds BU99006, is not the same as that which causes inhibition of MAO, and may point to the existence of another I(2) binding site.[1]


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