Age-associated insolubility of parkin in human midbrain is linked to redox balance and sequestration of reactive dopamine metabolites

• Published in: Acta neuropathologica Vol. 141; no. 5; pp. 725 - 754
• Main Authors: Tokarew, Jacqueline M., El-Kodsi, Daniel N., Lengacher, Nathalie A., Fehr, Travis K., Nguyen, Angela P., Shutinoski, Bojan, O’Nuallain, Brian, Jin, Ming, Khan, Jasmine M., Ng, Andy C. H., Li, Juan, Jiang, Qiubo, Zhang, Mei, Wang, Liqun, Sengupta, Rajib, Barber, Kathryn R., Tran, An, Im, Doo Soon, Callaghan, Steve, Park, David S., Zandee, Stephanie, Dong, Xiajun, Scherzer, Clemens R., Prat, Alexandre, Tsai, Eve C., Takanashi, Masashi, Hattori, Nobutaka, Chan, Jennifer A., Zecca, Luigi, West, Andrew B., Holmgren, Arne, Puente, Lawrence, Shaw, Gary S., Toth, Gergely, Woulfe, John M., Taylor, Peggy, Tomlinson, Julianna J., Schlossmacher, Michael G.
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.05.2021; Springer; Springer Nature B.V
• Subjects: Aging; Brain; Brain diseases; CD63 antigen; Dopamine; Dopamine receptors; Epitopes; Ethylenediaminetetraacetic acid; Free radicals (Chemistry); Hydrogen peroxide; Lysosomes; Medicin och hälsovetenskap; Medicine; Medicine & Public Health; Melanin; Mesencephalon; Metabolites; Monoclonal antibodies; Movement disorders; MPTP; Mutants; Neurodegeneration; Neurodegenerative diseases; Neurosciences; Original Paper; Oxidation; Oxidative stress; Parkin protein; Parkinson's disease; Pathology; Phenols; Redox reactions; Substantia nigra; Toxicity; Parkinsonism; Dopamine metabolism; gene; Anti-oxidant; Young-onset Parkinson disease; Redox chemistry; Neuromelanin; Parkin; PRKN/PARK2 gene
• ISSN: 0001-6322; 1432-0533; 1432-0533
• DOI: 10.1007/s00401-021-02285-4

The mechanisms by which parkin protects the adult human brain from Parkinson disease remain incompletely understood. We hypothesized that parkin cysteines participate in redox reactions and that these are reflected in its posttranslational modifications. We found that in post mortem human brain, including in the Substantia nigra , parkin is largely insoluble after age 40 years; this transition is linked to its oxidation, such as at residues Cys95 and Cys253. In mice, oxidative stress induces posttranslational modifications of parkin cysteines that lower its solubility in vivo. Similarly, oxidation of recombinant parkin by hydrogen peroxide (H 2 O 2 ) promotes its insolubility and aggregate formation, and in exchange leads to the reduction of H 2 O 2 . This thiol-based redox activity is diminished by parkin point mutants, e.g., p.C431F and p.G328E. In  prkn- null mice, H 2 O 2  levels are increased under oxidative stress conditions, such as acutely by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine toxin exposure or chronically due to a second, genetic hit; H 2 O 2  levels are also significantly increased in parkin-deficient human brain. In dopamine toxicity studies, wild-type parkin, but not disease-linked mutants, protects human dopaminergic cells, in part through lowering H 2 O 2 . Parkin also neutralizes reactive, electrophilic dopamine metabolites via adduct formation, which occurs foremost at the primate-specific residue Cys95. Further, wild-type but not p.C95A-mutant parkin augments melanin formation in vitro. By probing sections of adult, human midbrain from control individuals with epitope-mapped, monoclonal antibodies, we found specific and robust parkin reactivity that co-localizes with neuromelanin pigment, frequently within LAMP-3/CD63 + lysosomes. We conclude that oxidative modifications of parkin cysteines are associated with protective outcomes, which include the reduction of H 2 O 2 , conjugation of reactive dopamine metabolites, sequestration of radicals within insoluble aggregates, and increased melanin formation. The loss of these complementary redox effects may augment oxidative stress during ageing in dopamine-producing cells of mutant  PRKN  allele carriers, thereby enhancing the risk of Parkinson’s-linked neurodegeneration.