title: Preparation and application of monoclonal antibodies against oxidized DJ-1. Significant elevation of oxidized DJ-1 in erythrocytes of early-stage Parkinson disease patients.
abstract: DJ-1 was initially identified as a novel oncogene and has recently been found to be a causative gene for a familial form of Parkinson's disease (PD), viz, PARK7. Cysteine residue at position 106 (Cys-106) in DJ-1 was found to be oxidized preferentially under oxidative stress. In the present study, we developed specific antibodies against Cys-106-oxidized DJ-1 using baculovirus particles displaying the surface glycoprotein gp64-fusion protein as the immunizing agent. Western blot analysis combined with two-dimensional gel electrophoresis revealed that these antibodies specifically recognized oxidized DJ-1. Furthermore, we developed a competitive enzyme-linked immunosorbent assay (ELISA) for detecting oxidized DJ-1 and measured blood levels of oxidized DJ-1 in PD patients (n=15). It was observed that the levels of oxidized DJ-1 in erythrocytes of unmedicated PD patients were markedly higher without overlap than those of medicated PD patients and healthy subjects. No significant difference was observed in DJ-1 levels between mediated and unmediated PD patient. These results suggest the oxidative modification of DJ-1 in PD patients and the potential application of the antibody for diagnosis of PD at early-stage.
authors.affiliation: Health Technology Research Center, National Institute of Advanced Industrial Science and Technology, Ikeda, Osaka, Japan. ysaito@mail.doshisha.ac.jp
authors.forename: Yoshiro
authors.initials: Y
authors.lastname: Saito
authors.forename: Takao
authors.initials: T
authors.lastname: Hamakubo
authors.forename: Yasukazu
authors.initials: Y
authors.lastname: Yoshida
authors.forename: Yoko
authors.initials: Y
authors.lastname: Ogawa
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Hara
authors.forename: Harutoshi
authors.initials: H
authors.lastname: Fujimura
authors.forename: Yasuharu
authors.initials: Y
authors.lastname: Imai
authors.forename: Hiroko
authors.initials: H
authors.lastname: Iwanari
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Mochizuki
authors.forename: Mototada
authors.initials: M
authors.lastname: Shichiri
authors.forename: Keiko
authors.initials: K
authors.lastname: Nishio
authors.forename: Tomoya
authors.initials: T
authors.lastname: Kinumi
authors.forename: Noriko
authors.initials: N
authors.lastname: Noguchi
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Kodama
authors.forename: Etsuo
authors.initials: E
authors.lastname: Niki
mesh_terms: D000368:Aged; D000704:Analysis of Variance; D000906:Antibodies / Q000378:metabolism*; D000978:Antiparkinson Agents / Q000494:pharmacology; D016367:Baculoviridae; D015153:Blotting, Western; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000378:metabolism*; D005260:Female; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D047908:Intracellular Signaling Peptides and Proteins / Q000097:blood / Q000276:immunology* / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D015513:Oncogene Proteins / Q000097:blood / Q000276:immunology* / Q000378:metabolism*; D010084:Oxidation-Reduction; D010300:Parkinson Disease / Q000097:blood / Q000188:drug therapy / Q000378:metabolism*; D000071617:Protein Deglycase DJ-1; D013997:Time Factors
chemical_list: D000906:Antibodies; D000978:Antiparkinson Agents; D047908:Intracellular Signaling Peptides and Proteins; D015513:Oncogene Proteins; D006861:Hydrogen Peroxide; C105131:PARK7 protein, human; D000071617:Protein Deglycase DJ-1