Author

Nouf Alyami

Date Approved

7-14-2015

Date Posted

6-22-2016

Degree Type

Open Access Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

Committee Member

Hedeel Evans, Ph.D., Chair

Committee Member

Deborah Heyl-Clegg, Ph.D.

Committee Member

Vance Kennedy, Ph.D.

Abstract

Aspartate transcarbamoylase (ATCase) and Dihydroorotase (DHOase) catalyze the second and third steps, respectively, in de novo pyrimidine biosynthesis. Both enzymes form an active complex (DAC) in Aquifex aeolicus, where loop A of DHOase interacts with a domain of ATCase. The main objective of this work is to determine the function of specific residues of loop A in DAC interactions and to alter the catalytic activities through disruption of the interface between the two enzymes from A. aeolicus. The ATCase and DHOase domains have been expressed in Escherichia coli and purified using affinity chromatography. The interface of the published three-dimensional structure of the non-covalently associated dodecamer of DHOase and ATCase from A. aeolicus was examined using bioinformatics, and a peptide was designed to block a specific hydrophobic region. In this work, several peptides have been synthesized by solid phase peptide synthesis (SPPS). Assays were conducted to determine the binding affinities of the peptides to the DHO-ATC complex.

Included in

Chemistry Commons

Share

COinS