Date of Graduation

Fall 2020

Degree

Master of Science in Chemistry

Department

Chemistry

Committee Chair

Natasha DeVore

Keywords

orphan cytochrome P450, enzyme, Bietti’s crystalline dystrophy, molecular biology, expression, purification, protein crystallography

Subject Categories

Biochemistry | Molecular Biology

Abstract

Within the Cytochrome P450 class of enzymes, there are a group known as the “orphan” cytochromes. The “orphan” classification comes from the poorly understood in vivo functionality and substrate specificity. Cytochrome P450 4V2 (CYP4V2) is one of these “orphans” and belongs to the CYP4 family. The CYP4 family is known for the omega oxidation of endogenous fatty acids. This family is most commonly found on chromosome 1 (CYP4ABXZ). CYP4V2 is unique in that its location is bound to chromosome 4 as discovered by Jiao in 2004. Mutations within the CYP4V2 gene have been associated with the disease known as Bietti’s Crystalline Dystrophy (BCD). BCD is an autosomal recessive disorder presenting with crystalline and lipid deposits in the cornea and retina. Gian Bietti first reported the disease in 1937 in three patients, and in 2004 Li and his research group identified the CYP4V2 gene to be the cause. The unknown nature surrounding wild-type CYP4V2 is where this research study gets its purpose. To start declassifying its “orphan” status, a method for the expression and purification of protein has been developed and optimized to obtain protein for functional characterization and structural biology studies. Preliminary binding data for wild-type enzyme has yet to be obtained, and a crystal hit has been identified with further optimization required.

Copyright

© Cody Lane Turner

Open Access

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