Our research focuses on proteins called lysine deacetylases (KDACs) or histone deacetylases (HDACs). KDACs chemically modify other cellular proteins, and they have been linked to a wide range of essential cellular behaviors. Aberrant KDAC activity appears to be linked to numerous disease states, including many cancers. However, because it is largely unknown which proteins are being modified by KDACs, the reasons for the links to diseases are unclear. Our goal is to understand the physical interactions between KDACs and other proteins. Approaches utilized in the lab include characterization of purified KDACs and genetic engineering to control KDAC behavior in cells. Greater understanding of these interactions, and identification of particular target proteins, may lead to new therapies and diagnostics for diseases via new cellular pathways and drug targets.
Current research areas include:
Approaches and techniques used include enzyme kinetics, molecular biology, gene expression analysis, protein spectroscopy, mass spectrometry, molecular modeling, cell culture, and genetic engineering.
Toro TB, Edenfield SA, Hylton BJ, Watt TJ. Chelatable trace zinc causes low, irreproducible KDAC8 activity. Anal. Biochem. 2018, 540-541, 9-14. PMID: 29100752
Toro TB, Painter RG, Haynes RA, Glotser EY, Bratton MR, Bryant JR, Nichols KA, Matthew-Onabanjo AN, Matthew AN, Bratcher DR, Perry CD, Watt TJ. Purification of metal-dependent lysine deacetylases with consistently high activity. Protein Exp. Purif. 2018, 141, 1-6. PMID: 28843507
Toro TB, Bryant JR, Watt TJ. Lysine deacetylases exhibit distinct changes in activity profiles due to fluorophore-conjugation of substrates. Biochemistry 2017, 56, 4549-4558. PMID: 28749131
Toro TB, Pingali S, Nguyen TP, Garrett DS, Dodson KA, Nichols KA, Haynes RA, Payton-Stewart F, Watt TJ. KDAC8 with high basal velocity is not activated by N-acetylthioureas. PLoS One 2016, 11, e0146900. PMID: 26745872
Toro TB, Watt TJ. KDAC8 substrate specificity quantified by a biologically-relevant, label-free deacetylation assay. Protein Sci. 2015, 24, 2020-2032. PMID: 26402585
Johanson KE, Watt TJ. Inquiry-based experiments for large-scale introduction to PCR and restriction enzyme digests. Biochem. Mol. Bio. Ed. 2015, 43, 441-448. PMID: 26503481
Toro TB, Nguyen TP, Watt TJ. An improved 96-well turbidity assay for T4 lysozyme activity. MethodsX 2015, 2, 256-262. PMID: 26150996
Johanson KE, Watt TJ, McIntyre NR, Thompson M. Purification and characterization of enzymes from yeast: an extended undergraduate laboratory sequence for large classes. Biochem. Mol. Bio. Ed. 2013, 41, 251-261. PMID: 23868379