Author

Bunce

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.

Included in

Chemistry Commons

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