Date of Award
12-2005
Degree Name
Master of Science
Department
Chemistry
First Advisor
Dr. David L. Huffman
Second Advisor
Dr. David Reinhold
Third Advisor
Dr. Subra Muralidharan
Access Setting
Masters Thesis-Open Access
Abstract
Wilsons disease is an autosomal recessive disorder of copper metabolism which results from mutations in the gene encoding for Wilson disease protein (WDP). The cytosolic N-terminus of WDP comprises six metal binding domains each containing a conserved metal binding motif, GMXCXXC. The contribution of these metal binding domains to copper homeostasis is poorly understood. We have cloned, expressed and purified residues 485 to 633 corresponding to N-terminal metal-binding domains 5 and 6 of WDP (WD5-6). Two of the 3 disease causing mutations in the N-terminal metal binding domains of WDP occur one in each of these domains and the two domains have been shown to be essential for copper induced response, catalytic phosphorylation and other functions of WDP.
We found that WD5-6 binds 2 equivalents of copper, has a pl of 4.97 and behaves as a monomer in solution. We also showed by NMR titration that WD5-6 accepts copper from the fourth metal binding domain of WDP, Cu(l)WD4, and not from Cu(l)Atoxl , the copper chaperone that delivers copper to WDP. Solution NMR structure of apo-WD5-6 was elucidated and found to consist of 2 ferrodoxin-like folds that behaves as a unit in solution.
Recommended Citation
Achila, David, "Cloning, Overexpression, Purification and Characterization on N-Terminal Metal Binding Domains 5 and 6 of Wilsons Disease Protein" (2005). Masters Theses. 4459.
https://scholarworks.wmich.edu/masters_theses/4459