Isolation, Expression, Purification and Biochemical Characterization of Wilson Protein Metal Binding Domains 2 and 1-2

Author

Peter Kennedy Sande, Western Michigan University

Date of Award

4-2009

Degree Name

Master of Science

Department

Chemistry

First Advisor

Dr. David L. Huffman

Second Advisor

Dr. Ekkehard Sinn

Third Advisor

Dr. David Reinhold

Access Setting

Masters Thesis-Campus Only

Abstract

Wilson protein is a P_IB-type ATPase that translocates copper to the trans- Gogli network for incorporation into ceruloplasmin. It also helps to excrete excess copper in the bile by transporting excess copper to vesicles; the cell removes the vesicles filled with copper by exocytosis. Mutations in the Wilson protein gene ATP7B lead to Wilson disease. The N-terminus of the Wilson protein has 6 metal binding domains that play a significant role in copper transfer in the cell. Wilson protein metal binding domains 2 and 1-2 were cloned, expressed, purified and biochemically characterized to help determine their role in the copper transfer process in the cell. The metallochaperone CCS domain 1, HAH1, its mutants HAH1C11A and HAH1C14A were studied to determine and understand their role in transport of copper to the Wilson protein N-terminal metal binding domains.

Recommended Citation

Sande, Peter Kennedy, "Isolation, Expression, Purification and Biochemical Characterization of Wilson Protein Metal Binding Domains 2 and 1-2" (2009). Masters Theses. 230.
https://scholarworks.wmich.edu/masters_theses/230