Mutation and oxidation of Cys46 and Cys53 allosterically modulate the glyoxalase and esterase activities of DJ-1.

DJ-1, also known as PARK7 (Parkinson's disease protein 7), is an oncoprotein and a causative factor for hereditary recessive Parkinson's disease. DJ-1 contains three cysteines Cys46 (C46), Cys53 (C53), and Cys106 (C106) undergoing oxidation modifications under oxidative stress and affecting the anti-oxidative role of DJ-1. While C106 is known as a key catalytic active site for the enzymatic activity of DJ-1, the roles of C46 and C53 in the enzymatic activity regulation, along with the underlying mechanisms, remain poorly understood. Here, we revealed that mutations of the two cysteines resulted in decrease of both the glyoxalase and esterase activities of DJ-1. Structural analyses manifested that C46 and C53 mutations caused allosteric effects on the conformations of the catalytic active sites, which subsequently altered the interactions with the substrates. Furthermore, through modulating the reducing agent in the DJ-1 solution, we obtained the samples with varying degrees of oxidation at C46 and C53, and evidenced the influence of the oxidation states of C46 and C53 on the enzymatic activity of DJ-1. Importantly, an enhanced enzymatic activity associated with the increased oxidation of C46 in a C53-mutated background compared to wild-type DJ-1 was found. These results provide insights into the role and underlying mechanism of the mutation and oxidation modification of C46 and C53 in the glyoxalase and esterase activity of DJ-1, and offer informative clues for developing allosteric drugs potentially improving the enzymatic activity of DJ-1, which may facilitate the treatment of Parkinson's disease.