Thesis

Protein microarrays reveal tert-butylhydroquinone interactions with enzymes

Quinones are a class of molecules whose effect on cell growth and apoptosis pathways are poorly understood, yet several quinones are widely used as food preservatives and in cosmetics. This study takes advantage of protein microarray technology to identify specific protein-quinone interactions and to establish which class of proteins that quinones commonly interact with. This will enable us to gain a greater understanding of the biological pathways quinones may influence. Protein microarrays are sophisticated biological tools that allow the simultaneous examination of vast numbers of protein interactions. In this study protein microarray technology was used to examine the interaction oftertbutylhydroquinone (tBHQ) with ~5000 yeast and ~3000 human proteins. Strong interactions identified using the protein arrays were subsequently validated using traditional biochemical assays. The results indicated that tBHQ has a tendency to interact with DNA binding proteins, especially kinases, and exhibits a non-specific, weak kinase inhibitory effect. The results of this study indicated that tBHQ (1) reduced the growth rate of yeast in culture, (2) positively interacted with the yeast, Sir2 protein, and (3) it specifically inhibited the deacetylase activity of human Sir2. Keywords: Microarray, tert-butylhydroquinone, quinones, SIR2, Antioxidant Response Element

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