Assessing the role of zinc in Parkin activity
Abstract/Contents
- Abstract
- Neurodegeneration, the progressive loss of structure and function of neurons in the human brain, results in several disorders such as Alzheimer disease, Parkinson disease, amyotropic lateral sclerosis, and Huntington disease. Among older adults, Parkinson disorder (PD) ranks as the second most common age-related neurodegenerative disorder. PD is caused by the loss of dopaminergic neurons, key cells in the motor system. The four main symptoms of PD are tremors, bradykinesia, rigidity, and impaired balance. To this date there are only medications that target specific symptoms -- none target the underlying pathophysiology. Most PD cases, ~90%, are sporadic with a typical age of onset at 60. However, certain forms of PD have been linked to genetic causes. One of the most studied familial forms of PD is autosomal recessive juvenile parkinsonism (ARJP). Approximately 50% of all forms of ARJP are linked to the mutations of the PARK2 gene, which produces the protein parkin. Parkin is an E3 ligase, which plays an important role in targeting protein substrates for degradation or modification through ubiquitin tagging. Parkin belongs to a class of RING (Really Interesting New Gene) proteins designated in the RBR (RING-between-RING) family of E3 ligases. RING domains are rich in cysteines and have a strong affinity for binding zinc ions. Recent studies of the RING domains in parkin suggest that binding of zinc may play an important role in maintaining stability and solubility, which in turn may regulate protein function. The goal of my project is to determine a mechanism of Parkin activity independent of PINK1. My results suggest that autoubiquitination of Parkin is affected by zinc and that regulation occurs within the picomolar range of free zinc concentrations in PINK1 independent assays. Additionally, E1-ubiquitin conjugation is affected by zinc and the inhibition of conjugation occurs in the nanomolar zinc levels. As an initial test of this hypothesis I examined Parkin's effects on mitophagy when the intracellular zinc concentration was changed or by mutation of a zinc binding site in Parkin. External zinc was applied in in vitro assays with a chelator to regulate levels. Using site directed mutagenesis I tested whether known zinc sites were required for the zinc mediated inhibition of Parkin.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2019; ©2019 |
| Publication date | 2019; 2019 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Javelosa, Edritz |
|---|---|
| Degree supervisor | Wang, Xinnan |
| Thesis advisor | Wang, Xinnan |
| Thesis advisor | Clandinin, Thomas R. (Thomas Robert), 1970- |
| Thesis advisor | Garner, Craig |
| Thesis advisor | Reimer, Richard J |
| Thesis advisor | Weis, William I |
| Degree committee member | Clandinin, Thomas R. (Thomas Robert), 1970- |
| Degree committee member | Garner, Craig |
| Degree committee member | Reimer, Richard J |
| Degree committee member | Weis, William I |
| Associated with | Stanford University, Neurosciences Program. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Edritz P. Javelosa. |
|---|---|
| Note | Submitted to the Neurosciences Program. |
| Thesis | Thesis Ph.D. Stanford University 2019. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2019 by Edritz Javelosa
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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