Date of Award

2012

Document Type

Thesis

Degree Name

Bachelors

Department

Natural Sciences

First Advisor

Walstrom, Katherine

Keywords

c. elegans, Kinetics, Biochemistry

Area of Concentration

Biochemistry

Abstract

Glyceraldehyde-3-phosphate dehydrogenase (GPD) is an enzyme which converts glyceraldehyde-3-phosphate (G3P) into 1,3-bisphosphoglycerate using NAD+ and Pi as substrates. Due to the involvement of GAPDH in glycolysis, it can be found almost universally among organisms. C. elegans contains 4 GPD genes, and these proteins have not been characterized kinetically. GPD-3 was chosen for this study due to its involvement in longevity in adult worms. The gpd-3 gene was inserted into the vector pTYB11, and the GPD-3 protein was overexpressed in E. coli cells. The GPD-3 purification was unsuccessful due to inefficient cleavage of the purification tag, so a preparation of endogenous GPD-2/GPD-3 was kinetically characterized. A Km of 0.3 mM and ~1 mM was obtained for NAD+ and G3P, respectively. These Km values are significantly higher than in other species studied, and suggest the potential for further analysis.

Rights

This bibliographic record is available under the Creative Commons CC0 public domain dedication. The New College of Florida, as creator of this bibliographic record, has waived all rights to it worldwide under copyright law, including all related and neighboring rights, to the extent allowed by law.

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