Biophysical Characterization of the N-Terminal Metal Binding Domains 4-6, 5-6 and Two Disease-Causing Mutations of the Human Wilson Protein

Author

Nastaran Salehi Marzijarani

Date of Award

8-2010

Degree Name

Master of Science

Department

Chemistry

First Advisor

Dr. David L. Huffman

Second Advisor

Dr. Ramakrishna Guda

Third Advisor

Dr. Sherine Obare

Access Setting

Masters Thesis-Campus Only

Abstract

Copper is an essential element in all living organisms, serving as a cofactor for many important proteins and enzymes. However it is extremely toxic when present in excessive amounts. Wilson disease is an autosomal recessive disorder of copper metabolism. The Wilson disease protein is a copper-transporting P_1b-type ATPase, ATP7B. Mutations in the Wilson disease protein lead to an accumulation of toxic levels of copper in the liver, brain, cornea and kidney, causing the hepatic and/or neurological symptoms accompanying this disease. The Wilson protein contains six N-terminal metal binding domains. Metal binding domains of ATP7B (WLN4-6, WLN5-6) and two disease-causing mutants (WLN5-6 V536A, WLN5-6 L492S) were cloned, expressed, and purified. Native protein and mutants were characterized by Gel Filtration, Dynamic Light Scattering, Circular Dichroism, and Fluorescence spectroscopy (steady-state and time-resolved methods). It was concluded that the disease-causing mutations V536A and L492S significantly perturb the biophysical properties of WLN5-6.

Recommended Citation

Marzijarani, Nastaran Salehi, "Biophysical Characterization of the N-Terminal Metal Binding Domains 4-6, 5-6 and Two Disease-Causing Mutations of the Human Wilson Protein" (2010). Masters Theses. 359.
https://scholarworks.wmich.edu/masters_theses/359