The objective of this research program is to elucidate, at the molecular level, the mechanism and function of copper and zinc binding in the prion protein (PrP) and the relationship of these interactions to neuron maintenance and neurological disease. A misfolded form of the prion protein (PrP) is responsible for a class of fatal neurodegenerative diseases termed the Transmissible Spongiform Encephalopathies (TSEs), which include mad cow disease and Creutzfeldt-Jakob disease in humans. PrP is a highly abundant, membrane-bound glycoprotein found in all mammals and avian species. Although originally recognized for its role in neurodegenerative disease, it is now clear that PrP is essential for sustaining neuron function and regulating apoptosis. PrP takes up both copper and zinc at sites are essential for PrP activity. Over the last funding period, the PI's laboratory characterized structural and thermodynamic features of these metal ion sites and also identified a remarkable link between certain mutations and onset age in inherited prion disease. The next funding period will build on these findings using both magnetic resonance and cellular studies with the following projects: 1) determine the structure of the full-length, copper or zinc occupied protein;2) evaluate how metal ions influence in vivo cleavage, implicated in apoptosis regulation;and 3) examine how structure and cellular behavior is altered by mutations associated with inherited disease, and also by mutations that prevent disease. Moreover, project three will apply new synthetic strategies for investigating the role of protein glycosylation. It is anticipated that these studies will provide fundamental new concepts for understanding the role of both PrP and copper in basic neurochemistry, and strategies for treating neurodegenerative disease and brain trauma.

Public Health Relevance

This research is aimed at evaluating proteins of the central nervous system that regulate copper and other micronutrients. These proteins are essential for neuron health and understanding their behavior will lead to new strategies for treating neurodegenerative diseases, such as Alzheimer's, and treating brain trauma.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM065790-10
Application #
8248205
Study Section
Macromolecular Structure and Function A Study Section (MSFA)
Program Officer
Anderson, Vernon
Project Start
2002-03-01
Project End
2015-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
10
Fiscal Year
2012
Total Cost
$278,484
Indirect Cost
$87,484
Name
University of California Santa Cruz
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
125084723
City
Santa Cruz
State
CA
Country
United States
Zip Code
95064
Evans, Eric G B; Pushie, M Jake; Markham, Kate A et al. (2016) Interaction between Prion Protein's Copper-Bound Octarepeat Domain and a Charged C-Terminal Pocket Suggests a Mechanism for N-Terminal Regulation. Structure 24:1057-67
Lau, Agnes; McDonald, Alex; Daude, Nathalie et al. (2015) Octarepeat region flexibility impacts prion function, endoproteolysis and disease manifestation. EMBO Mol Med 7:339-56
Evans, Eric G B; Millhauser, Glenn L (2015) Genetic Incorporation of the Unnatural Amino Acid p-Acetyl Phenylalanine into Proteins for Site-Directed Spin Labeling. Methods Enzymol 563:503-27
Zhang, Siyuan; Naab, Benjamin D; Jucov, Evgheni V et al. (2015) n-Dopants Based on Dimers of Benzimidazoline Radicals: Structures and Mechanism of Redox Reactions. Chemistry 21:10878-85
Stellato, Francesco; Minicozzi, Velia; Millhauser, Glenn L et al. (2014) Copper-zinc cross-modulation in prion protein binding. Eur Biophys J 43:631-42
McDonald, Alex J; Millhauser, Glenn L (2014) PrP overdrive: does inhibition of α-cleavage contribute to PrP(C) toxicity and prion disease? Prion 8:
McDonald, Alex J; Dibble, Jessie P; Evans, Eric G B et al. (2014) A new paradigm for enzymatic control of *-cleavage and *-cleavage of the prion protein. J Biol Chem 289:803-13
Thompson, Darren A; Evans, Eric G B; Kasza, Tomas et al. (2014) Adapter reagents for protein site specific dye labeling. Biopolymers 102:273-9
Pushie, M Jake; Nienaber, Kurt H; McDonald, Alex et al. (2014) Combined EXAFS and DFT structure calculations provide structural insights into the 1:1 multi-histidine complexes of Cu(II) , Cu(I) , and Zn(II) with the tandem octarepeats of the mammalian prion protein. Chemistry 20:9770-83
McDonald, Alex; Pushie, M Jake; Millhauser, Glenn L et al. (2013) New insights into metal interactions with the prion protein: EXAFS analysis and structure calculations of copper binding to a single octarepeat from the prion protein. J Phys Chem B 117:13822-41

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