Distal end of 105–125 loop – A putative reductase binding domain of phthalate dioxygenase

Abstract

The phthalate dioxygenase system consists of the dioxygenase, PDO, which contains a Rieske [2Fe–2S] center and a Fe(II)-mononuclear center, and the reductase, PDR. Involvement of the distal end of the 105–125 loop of PDO in its interaction with PDR was tested by substituting charged residues in the loop with alanines and by replacing the conserved tryptophan-94. Compared to wild-type PDO, all variants had lower catalytic activity and the Rieske centers were reduced more slowly by reduced PDR. The rates of oxidation of the Rieske centers by oxygen, which represent electron transfer between the Rieske and mononuclear centers, were essentially unaffected. These results suggest that positively charged residues of the distal end of the 105–125 loop are collectively involved in PDR binding with the PDO. Contrary to expectations, Trp94 variants were not directly involved in electron transfer between PDR and PDO. The tryptophan appears to have mainly a structural role, apparently preserving the hydrophilic environment of the Rieske center.