Novel Redox-Dependent Esterase Activity (EC 3.1.1.2) for DJ-1: Implications for Parkinson's Disease
Loading...
Authors
Vazquez-Mayorga, Emmanuel
Diaz-Sanchez, Angel G.
Dagda, Ruben K.
Dominguez-Solis, Carlos A.
Dagda, Raul Y.
Coronado-Ramirez, Cynthia K.
Martinez-Martinez, Alejandro
Issue Date
2016
Type
Article
Language
Keywords
4-nitrophenyl acetate , human carboxyl esterase , redox sensor , oxidative stress , DJ1/PARK7 , ROS , Parkinson's disease
Alternative Title
Abstract
Mutations the in human DJ-1 (hDJ-1) gene are associated with early-onset autosomal recessive forms of Parkinson's disease (PD). hDJ-1/parkinsonism associated deglycase (PARK7) is a cytoprotective multi-functional protein that contains a conserved cysteine-protease domain. Given that cysteine-proteases can act on both amide and ester substrates, we surmised that hDJ-1 possessed cysteine-mediated esterase activity. To test this hypothesis, hDJ-1 was overexpressed, purified and tested for activity towards 4-nitrophenyl acetate (pNPA) as mu mol of pNPA hydrolyzed/min/mg . protein (U/mg protein). hDJ-1 showed maximum reaction velocity esterase activity (V-max = 235.10 +/- 12.00 U/mg protein), with a sigmoidal fit (S-0.5 = 0.55 +/- 0.040 mM) and apparent positive cooperativity (Hill coefficient of 2.05 +/- 0.28). A PD-associated mutant of DJ-1 (M26I) lacked activity. Unlike its protease activity which is inactivated by reactive oxygen species (ROS), esterase activity of hDJ-1 is enhanced upon exposure to low concentrations of hydrogen peroxide (< 10 mu M) and plateaus at elevated concentrations (> 100 mu M) suggesting that its activity is resistant to oxidative stress. Esterase activity of DJ-1 requires oxidation of catalytic cysteines, as chemically protecting cysteines blocked its activity whereas an oxido-mimetic mutant of DJ-1 (C106D) exhibited robust esterase activity. Molecular docking studies suggest that C106 and L126 within its catalytic site interact with esterase substrates. Overall, our data show that hDJ-1 contains intrinsic redox-sensitive esterase activity that is abolished in a PD-associated mutant form of the hDJ-1 protein.
Description
Citation
Vázquez-Mayorga, E., Díaz-Sánchez, Á., Dagda, R., Domínguez-Solís, C., Dagda, R., Coronado-Ramírez, C., & Martínez-Martínez, A. (2016). Novel Redox-Dependent Esterase Activity (EC 3.1.1.2) for DJ-1: Implications for Parkinson's Disease. International Journal of Molecular Sciences, 17(8), 1346. doi:10.3390/ijms17081346
Publisher
License
Creative Commons Attribution 4.0 International
Journal
Volume
Issue
PubMed ID
ISSN
1422-0067