Date of Award

4-1-2022

Document Type

Honors Thesis

Department

Biology

First Advisor

Peter Lyons

Abstract

Carboxypeptidase A1 (CPA1) is an enzyme utilized in digestion. CPA1 contains a prodomain region, which must be cleaved for enzymatic function. The prodomain region has been hypothesized to be necessary for structural stability, as previous studies were unable to produce a functional CPA1 protein without the prodomain region. However, the recent discovery of a related enzyme, Carboxypeptidase 0 (CP0), which does not have a prodomain yet functions enzymatically, challenges this assertion. To determine if the prodomain is essential to the folding of CPA1, an HA-tag was added for detection, and the prodomain was deleted via PCR. A Western blot was performed to determine expression and enzyme assays were conducted to determine function. CPA1 without a prodomain was detected on the Western blot, but did not display activity in the enzyme assays. This indicates that CPA1 without a prodomain folds adequately enough to be expressed but is nonfunctional. Therefore, the prodomain region appears to be necessary for CPA1 structure and functionality.

Subject Area

Carboxypeptidases; Enzymes; Digestive enzymes

Included in

Biology Commons

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