Recently,alpha-synuclein has been identified as a major component of Lewy bodies, the intracellular inclusions that are a pathological hallmark of Parkinson's disease (PD). Our goals in this proposal are to test the hypothesis that a critical step in Parkinson's disease is the aggregation of alpha-synuclein, which leads to the formation of Lewy Bodies and subsequently to neuronal death. Specifically we will determine the molecular basis for alpha-synuclein aggregation and investigate potential inhibitors of alpha-syouclein aggregation. Our preliminary results have revealed a number of factors that lead to a confonnational change in alpha synuclein at neutral pH, and also to aggregation and fibril formation. We plan a systematic characterization of the biophysical properties of alpha-synuclein to determine if there is a correlation between its conformation and its propensity to aggregate, with both wild type and mutant alpha-synucleins. We will investigate whether various factors associated with PD, for example, metal ions and pesticides, enhance the aggregation of alpha-synuclein. Details of the aggregation process will be studied to elucidate the molecular mechanism of aggregation and fibril formation. We will screen a series of peptides and small molecules for inhibitory effects on alpha-synuclein aggregation. These experiments represent critical steps towards elucidating the role of alpha-synuclein in Parkinson's disease. We expect to learn the potential role of various factors, ranging from environmental contaminants to the concentration of alpha-synuclein, in triggering fibril formation. Further, we anticipate fnding inhibitors which may lay the groundwork for potential therapeutic approaches. Techniques to be used include various biophysical/biochemical methods, such as attenuated total reflectance FTIR to analyze the conformaffonal state of aggregated alpha-synuclein, atomic force and electron microscopy to image the aggregates, and kinetic methods to monitor the rate of formation of fibrils.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Project (R01)
Project #
5R01NS039985-02
Application #
6394360
Study Section
Special Emphasis Panel (ZRG1-MDCN-2 (01))
Program Officer
Murphy, Diane
Project Start
2000-04-10
Project End
2003-03-31
Budget Start
2001-04-01
Budget End
2002-03-31
Support Year
2
Fiscal Year
2001
Total Cost
$285,138
Indirect Cost
Name
University of California Santa Cruz
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
City
Santa Cruz
State
CA
Country
United States
Zip Code
95064
Hong, Dong-Pyo; Han, Shubo; Fink, Anthony L et al. (2011) Characterization of the non-fibrillar ýý-synuclein oligomers. Protein Pept Lett 18:230-40
Zhou, Wenbo; Long, Chunmei; Reaney, Stephen H et al. (2010) Methionine oxidation stabilizes non-toxic oligomers of alpha-synuclein through strengthening the auto-inhibitory intra-molecular long-range interactions. Biochim Biophys Acta 1802:322-30
Zhou, Wenbo; Gallagher, Amy; Hong, Dong-Pyo et al. (2009) At low concentrations, 3,4-dihydroxyphenylacetic acid (DOPAC) binds non-covalently to alpha-synuclein and prevents its fibrillation. J Mol Biol 388:597-610
Meng, Xiaoyun; Munishkina, Larissa A; Fink, Anthony L et al. (2009) Molecular mechanisms underlying the flavonoid-induced inhibition of alpha-synuclein fibrillation. Biochemistry 48:8206-24
Hong, Dong-Pyo; Fink, Anthony L; Uversky, Vladimir N (2009) Smoking and Parkinson's disease: does nicotine affect alpha-synuclein fibrillation? Biochim Biophys Acta 1794:282-90
Munishkina, L A; Fink, A L; Uversky, V N (2009) Accelerated fibrillation of alpha-synuclein induced by the combined action of macromolecular crowding and factors inducing partial folding. Curr Alzheimer Res 6:252-60
Hu, Dongmei; Qin, Zhijie; Xue, Bin et al. (2008) Effect of methionine oxidation on the structural properties, conformational stability, and aggregation of immunoglobulin light chain LEN. Biochemistry 47:8665-77
Hong, Dong-Pyo; Fink, Anthony L; Uversky, Vladimir N (2008) Structural characteristics of alpha-synuclein oligomers stabilized by the flavonoid baicalein. J Mol Biol 383:214-23
Munishkina, Larissa A; Fink, Anthony L; Uversky, Vladimir N (2008) Concerted action of metals and macromolecular crowding on the fibrillation of alpha-synuclein. Protein Pept Lett 15:1079-85
Munishkina, Larissa A; Ahmad, Atta; Fink, Anthony L et al. (2008) Guiding protein aggregation with macromolecular crowding. Biochemistry 47:8993-9006

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