Date of Award
8-2008
Degree Name
Master of Science
Department
Chemistry
First Advisor
Dr. David L. Huffman
Second Advisor
Dr. Michael Barcelona
Third Advisor
Dr. David Reinhold
Access Setting
Masters Thesis-Open Access
Abstract
Wilson disease is an autosomal recessive disorder resulting in the accumulation of copper in the body primarily in the liver and brain. In order to understand the pathogenesis of Wilson disease, our laboratory is studying several different proteins. Wilson disease protein (ATP7B) is a P-type ATPase which acts to remove excess copper from the cells. The p62 subunit of the dynactin complex is a protein whose function is still unknown but it has been proposed to have a copperdependent interaction with ATP7B. HAH1 is a human metallochaperone that binds copper as it comes into the cells from CTR1, a permease. HAH1 then shuttles copper ions to one or more of the domains at the N-terminus of ATP7B. Copper binding occurs via the thiol groups on two cysteines of HAH1 and the copper-binding domains of ATP7B.
In order to study protein-protein interactions we have cloned and expressed these proteins. Three different vector constructs of p62 protein were prepared and tested. Using site-directed mutagenesis we also generated mutants of both HAH1 and WLN4 (copper-binding domain 4 of ATP7B) and used them to study copperdependent interactions. We have used the two mutants pHAH1(C11A,C14A) and pTRX-WLN4(C15A,C18A) to serve as controls in the study of copper exchange between HAH1 and WLN4.
Recommended Citation
Oshule, Paul Sifuna, "Characterization of p62, HAH1 and their Interaction with the Wilson Disease Protein" (2008). Masters Theses. 4323.
https://scholarworks.wmich.edu/masters_theses/4323