title: Novel Redox-Dependent Esterase Activity (EC 3.1.1.2) for DJ-1: Implications for Parkinson's Disease.
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 µmol of pNPA hydrolyzed/min/mg·protein (U/mg protein). hDJ-1 showed maximum reaction velocity esterase activity (Vmax = 235.10 ± 12.00 U/mg protein), with a sigmoidal fit (S0.5 = 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 µM) and plateaus at elevated concentrations (>100 µM) 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.
authors.affiliation: Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo envolvente Pronaf y Estocolmo s/n, Ciudad Juarez, Chihuahua 32310, Mexico. emmanuel.vazquez@uacj.mx.|Department of Pharmacology, University of Nevada, Reno School of Medicine, Mailstop 318, Manville Building 19A(Office)/18(Lab), Reno, NV 89557, USA. emmanuel.vazquez@uacj.mx.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Vázquez-Mayorga
authors.affiliation: Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo envolvente Pronaf y Estocolmo s/n, Ciudad Juarez, Chihuahua 32310, Mexico. angel.diaz@uacj.mx.
authors.forename: Ángel G
authors.initials: ÁG
authors.lastname: Díaz-Sánchez
authors.affiliation: Department of Pharmacology, University of Nevada, Reno School of Medicine, Mailstop 318, Manville Building 19A(Office)/18(Lab), Reno, NV 89557, USA. rdagda@medicine.nevada.edu.
authors.forename: Ruben K
authors.initials: RK
authors.lastname: Dagda
authors.affiliation: Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo envolvente Pronaf y Estocolmo s/n, Ciudad Juarez, Chihuahua 32310, Mexico. al135792@alumnos.uacj.mx.
authors.forename: Carlos A
authors.initials: CA
authors.lastname: Domínguez-Solís
authors.affiliation: Department of Pharmacology, University of Nevada, Reno School of Medicine, Mailstop 318, Manville Building 19A(Office)/18(Lab), Reno, NV 89557, USA. rauld@medicine.nevada.edu.
authors.forename: Raul Y
authors.initials: RY
authors.lastname: Dagda
authors.affiliation: Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo envolvente Pronaf y Estocolmo s/n, Ciudad Juarez, Chihuahua 32310, Mexico. al113722@alumnos.uacj.mx.
authors.forename: Cynthia K
authors.initials: CK
authors.lastname: Coronado-Ramírez
authors.affiliation: Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo envolvente Pronaf y Estocolmo s/n, Ciudad Juarez, Chihuahua 32310, Mexico. alejandro.martinez@uacj.mx.|El Colegio de Chihuahua, Calle Partido Díaz 4723 esquina con Anillo Envolvente del Pronaf, colonia Progresista, Ciudad Juárez, Chihuahua 32310, Mexico. alejandro.martinez@uacj.mx.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Martínez-Martínez
mesh_terms: D004950:Esterases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D062105:Molecular Docking Simulation; D009154:Mutation; D009596:Nitrophenols / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D010300:Parkinson Disease / Q000201:enzymology* / Q000235:genetics; D000071617:Protein Deglycase DJ-1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D009596:Nitrophenols; D017382:Reactive Oxygen Species; C008642:4-nitrophenyl acetate; D006861:Hydrogen Peroxide; D004950:Esterases; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1
keywords: 4-nitrophenyl acetate; DJ1/PARK7; Parkinson’s disease; ROS; human carboxyl esterase; oxidative stress; redox sensor