1-Aminocyclopropane-1-carboxylic acid oxidase (ACCO) is a nonheme Fe(II)-containing enzyme that is related to the 2-oxoglutarate-dependent dioxygenase family. The binding of substrates/cofactors to tomato ACCO was investigated through kinetics, tryptophan fluorescence quenching, and modeling studies. α-Aminophosphonate analogs of the substrate (1-aminocyclopropane-1-carboxylic acid, ACC), 1-aminocyclopropane-1-phosphonic acid (ACP) and (1-amino-1-methyl)ethylphosphonic acid (AMEP), were found to be competitive inhibitors versus both ACC and bicarbonate (HCO(3)(-)) ions. The measured dissociation constants for Fe(II) and ACC clearly indicate that bicarbonate ions improve both Fe(II) and ACC binding, strongly suggesting a stabilization role for this cofactor. A structural model of tomato ACCO was constructed and used for docking experiments, providing a model of possible interactions of ACC, HCO(3)(-), and ascorbate at the active site. In this model, the ACC and bicarbonate binding sites are located close together in the active pocket. HCO(3)(-) is found at hydrogen-bond distance from ACC and interacts (hydrogen bonds or electrostatic interactions) with residues K158, R244, Y162, S246, and R300 of the enzyme. The position of ascorbate is also predicted away from ACC. Individually docked at the active site, the inhibitors ACP and AMEP were found coordinating the metal ion in place of ACC with the phosphonate groups interacting with K158 and R300, thus interlocking with both ACC and bicarbonate binding sites. In conclusion, HCO(3)(-) and ACC together occupy positions similar to the position of 2-oxoglutarate in related enzymes, and through a hydrogen bond HCO(3)(-) likely plays a major role in the stabilization of the substrate in the active pocket.

A mu-oxo-diiron(III) complex bridged by two molecules of 1-aminocyclopropane-1-carboxylic acid (ACCH) was prepared with the ligand 1,4,7-triazacyclononane (TACN): [(TACN)Fe(2)(mu-O)(mu-ACCH)(2)](ClO(4))(4) x 2 H(2)O (1). This complex was characterized, and its crystal structure was solved. The bridging amino acid moieties were found in their zwitterionic forms (noted as ACCH). Reactivity assays were performed in the presence of hydrogen peroxide, and 1 turned out to be the first example of a well-characterized iron-ACCH complex able to produce ethylene from the bound ACCH moiety. The reaction requires the presence of a few equivalents of base, probably involved in the deprotonation of the amine groups of the ACCH bridges.