Date of Award
4-2005
Degree Name
Master of Science
Department
Chemistry
First Advisor
Dr. David L. Huffman
Second Advisor
Dr. Michael J. Barcelona
Third Advisor
Dr. Marc Perkovic
Fourth Advisor
Dr. Brian Tripp
Access Setting
Masters Thesis-Open Access
Abstract
The human copper-transporting ATPase Wilson's disease protein was one of the first proteins shown to be actively involved in cellular copper homeostasis. The protein includes six N-terminal metal binding domains. The individual capabilities of the domains to bind and transfer copper are not yet fully understood. Each metal-binding domain is about 72 amino acid residues long and contains the conserved binding motif GMTCXXC, of which the cysteines are involved in the copper coordination. Studies have shown that the protein binds six copper atoms suggesting the potential involvement of each metal-binding domain. Separate functional roles for the latter domains have been proposed implicating a possible interaction between the domains.
Here the first and fourth domains have been isolated and characterized by several chemical means including high resolution gel filtration, MALDI mass spectroscopy, and isoelectric focusing. Each domain demonstrated copper binding capabilities. Copper exchange activity has demonstrated between the two individual metal-binding domains in vitro using varying molar ratios between donor and acceptor. The exchange event was determined to be reversible and equilibrium was quickly achieved in less than 5 minutes. The Kexchange constant is estimated at 0.69 for the reaction.
Recommended Citation
Bunce, Jennifer, "Copper Exchange Studies between the First and Fourth N-Terminal Metal-Binding Domains of Wilson Disease ATPase" (2005). Masters Theses. 4462.
https://scholarworks.wmich.edu/masters_theses/4462