Summary
The identification of monogenic variants of Parkinson’s disease (PD) has provided novel insights into its unknown pathogenesis. As the first protein linked to autosomal-recessive forms of PD, Parkin became a welcome tool to explain biochemical and neuropathological observations that had suggested involvement of the ubiquitin-proteasome system (UPS) in PD. Based on cellular expression studies and biochemical in vitro experiments, several researchers ascribed an E3-type, E2-dependent ubiquitin protein ligase activity to wild-type (but not mutant) Parkin proteins. Although the individual components of the proposed Parkin ubiquitin ligase complex in the normal human brain remain to be identified and the E3 ligase effect of Parkin function has not yet been confirmed in an animal model, the scientific exploration of a protein with several links to the UPS has provided many leads in PD research. This chapter describes assays that the authors have used to examine the cellular and in vitro effects of neural Parkin.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Jellinger, K. A. and Mizuno, Y. (2003) Parkinson’s disease. In Neurodegeneration: The Molecular Pathology of Dementia and Movement Disorders (Dickson, D. W., ed.), ISN Neuropath Press, Basel, pp. 159–186.
Dauer, W. and Prezdborski, S. (2003) Parkinson’s disease: mechanisms and models. Neuron 39, 889–909.
Chiechanover, A. and Brundin, P. (2003) The ubiquitin proteasome system in neurodegenerative diseases: sometimes the chicken, sometimes the egg. Neuron 40, 427–446.
Golbe, L. I. and Maroudian, M. M. (2004) Alpha-synuclein in Parkinson’s disease: light from two new angles. Annal. Neurol. 55, 153–155.
Singleton, A., Farrer, M., Johnson J., et al. (2003) Alpha-synuclein locus triplication causes Parkinson’s disease. Science 302, 841.
Kitada, T., Asakawa, S., Hattori, N., et al. (1998) Mutations in the parkin gene cause autosomal-recessive juvenile parkinsonism. Nature 392, 605–608.
Oliveira, S. A., Scott, W. K., Martin, E. R., et al. (2003) Parkin mutations and susceptibility alleles in late-onset Parkinson’s disease. Ann. Neurol. 53, 624–629.
Hasegawa, M., Fujiwara, H., Nonaka, T., et al. (2002) Phosphorylated alpha-synuclein is ubiquitinated in alpha-synucleinopathy lesions. J. Biol. Chem. 277, 49071–49076.
Schlossmacher, M. G., Frosch, M. P., Gai, W. P., et al. (2002) Parkin localizes to the Lewy bodies of Parkinson disease and dementia with Lewy bodies Am. J. Pathol. 160, 1655–1667.
Leroy, E., Boyer, R., Auburger, G., et al. (1998) The ubiquitin pathway in Parkinson’s disease. Nature 395, 451–452.
Liu, Y., Fallon, L., Lashuel, H. A., Liu, Z., and Lansbury, P. T. (2002) The UCH-L1 gene encodes two opposing enzymatic activities that affect alpha-synuclein degradation and Parkinson’s disease susceptibility. Cell 111, 209–218.
Shimura, H., Hattori, N., Kubo, S., et al. (2000) Familial Parkinson disease gene product, parkin, is a ubiquitin-protein ligase Nat. Gen. 25, 302–305.
Zhang, Y., Gao, J., Chung, K. K., et al. (2000) Parkin functions as an E2-dependent ubiquitin-protein ligase and promotes the degradation of the synaptic vesicle-associated protein, CDCrel-1. Proc. Nat. Acad. Sci. USA 97, 13354–13359.
Imai, Y., Soda, M., and Takahashi, R. (2000) Parkin suppresses unfolded protein stress-induced cell death through its E3 ubiquitin-protein ligase activity. J. Biol. Chem. 275, 35661–35664.
Lücking, C. B., Durr, A., Bonifati, V., et al. (2000) Association between early-onset Parkinson’s disease and mutations in the parkin gene. French Parkinson’s disease genetics study group. N. Engl. J. Med. 342, 1560–1567.
Goldberg, M. S., Fleming, S. M., Palacino, J. J., et al. (2003) Parkin-deficient mice exhibit nigrostriatal deficits but no loss of dopaminergic neurons. J. Biol. Chem. 278, 43628–43635.
Itier, J. M., Ibanez, P., Mena, M. A., et al. (2003) Parkin gene inactivation alters behaviour and dopamine neurotransmission in the mouse. Hum. Mol. Gen. 12, 2277–2291.
Shimura, H., Hattori, N., Kubo, S., et al. (1999) Immunohistochemical and subcellular localization of parkin protein: absence of protein in autosomal recessive juvenile parkinsonism patients. Ann. Neurol. 45, 668–672.
Shimura, H., Schlossmacher, M. G., Hattori, N., et al. (2001) Ubiquitination of a new form of alpha-synuclein by parkin from human brain: implications for Parkinson’s disease. Science 293, 263–269.
Cookson, M. R., Lockhart, P. J., McLendon, C., O’Farrell, C., Schlossmacher, M., and Farrer, M. J. (2003) RING finger 1 mutations in parkin produce altered localization of the protein. Hum. Mol. Gen. 12, 2957–2965.
Petrucelli, L., O’Farrell, C., Lockhart, P. J., et al. (2003) Parkin protects against the toxicity associated with mutant alpha-synuclein: proteasome dysfunction selectively affects catecholaminergic neurons. Neuron 36, 1007–1019.
Staropoli, J. F., McDermott, C., Martinat, C., et al. (2003) Parkin is a component of an SCF-like ubiquitin ligase complex and protects postmitotic neurons from kainate toxicity. Neuron 37, 735–749.
Yang, Y., Nishimura, I., Imai, Y., Takahashi, R., and Lu, B. (2003) Parkin suppresses dopaminergic neuron-selective neurotoxicity by Pael-R in Drosophila. Neuron 37, 911–924.
Choi, P., Snyder, H., Petrucelli, H., et al. (2003) SEPT_v2 is a parkin-binding protein. Brain Res. Mol. Brain Res. 117, 179–189.
Kyte, J. and Doolittle, R. F. (1982) A simple method for displaying the hydropathic character of a protein J. Mol. Biol. 157, 105–132.
http://www.openbiosystems.com/custom_peptide_antibody_services.php .
Martinez, M. C., Ochiishi, T., Majewski, M., and Kosik, S. K. (2003) Dual regulation of neuronal morphogenesis by a delta-catenin-cortactin complex and Rho. J. Cell. Biol. 162, 99–111.
Stoltzner, S. E., Grenfell, T. J., Mori, C., Wisniewski, K. E., Selkoe, D. J., and Lemere, C. A. (2000) Temporal accrual of complement proteins in amyloid plaques in Down’s syndrome with Alzheimer’s disease. Am. J. Pathol. 156, 489–499.
Schlossmacher, M. G., Cullen, V., and Müthing, J. (2005) Glucocerebrosidase, α-synuclein and Parkinson’s disease. New Engl. J. Med., in press.
Acknowledgments
The authors thank M. Baker for advice regarding antibody production, M. P. Frosch and J. A. Chan for human brain tissue collection, C. Lemere for sharing her immunohistochemical protocol, N. Hattori and Y. Mizuno for the human parkin cDNA, M. Medina for the generation of stably transfected SY5Y cells, and K. S. Rasakham and P. Xu for technical assistance. This work was supported by the Grass Foundation (Braintree, MA), the Lefler Foundation (Harvard Medical School), the M.S.A. Fund (Brigham & Women’s Hospital), and the NINDS-NIH (NS02127) to M. Schlossmacher and a Pergolide Fellowship (Eli Lilly, Japan K. K.) to H. Shimura.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2005 Humana Press Inc.
About this protocol
Cite this protocol
Schlossmacher, M.G., Shimura, H. (2005). Parkinson’s Disease. In: Patterson, C., Cyr, D.M. (eds) Ubiquitin-Proteasome Protocols. Methods in Molecular Biology™, vol 301. Humana Press. https://doi.org/10.1385/1-59259-895-1:351
Download citation
DOI: https://doi.org/10.1385/1-59259-895-1:351
Publisher Name: Humana Press
Print ISBN: 978-1-58829-252-0
Online ISBN: 978-1-59259-895-3
eBook Packages: Springer Protocols