Author

Ochieng

Date of Award

4-2007

Degree Name

Master of Science

Department

Chemistry

First Advisor

Dr. David L. Huffman

Second Advisor

Dr. Sherine Obare

Third Advisor

Dr. Dongil Lee

Access Setting

Masters Thesis-Open Access

Abstract

Wilson protein (ATP7B) is a copper-transporting P-type ATPase that regulates copper homeostasis and secretion of copper-containing enzymes in the liver. In hepatocytes, ATP7B delivers copper to apoceruloplasmin and mediates the excretion of excess copper into bile. Mutation in the gene coding for Wilson protein leads to Wilson disease. Several mutations in Wilson protein have been identified and of particular interest, mutations affecting the N-metal binding region significantly interfere with the ability of Wilson protein to transport copper to its target in the copper-trafficking pathway. In an attempt to characterize the disease-causing mutation (G591D) in Wilson protein domain 6 (WLN6), we have sub-cloned, overexpressed, purified and structurally characterized the native domain 6 construct of Wilson protein using NMR. We have also expressed the G591D mutant form of the same domain in GST fusion for comparison studies. Solution NMR results indicate that native WNL6 is monomeric and well-folded with a βαββαβ ferroxidin-like structure. Subsequent structural determination of this mutant with NMR will reveal more about the effect of this mutation.

Included in

Chemistry Commons

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