Catechol-O-methyltransferase (COMT) is inhibited rapidly and irreversibly by N-(3,4-dihydroxyphenyl) maleimide. S-adenosylmethionine (AdoMet) and magnesium ions protect the enzyme from inactivation by this compound, but no protection is observed by the catechol substrate. However, the corresponding succinimide analogue shows a reversible inhibition of COMT, which is competitive with pyrocatecholphthalein and non-competitive with AdoMet. Amino-group reagents also inhibit COMT and this inhibition is protected by AdoMet, suggesting that sulphydryl and amino groups essential for activity are located in an AdoMet-binding site on COMT. The maleimide derivative may be considered to be an active-site directed inhibitor.