The objective of the proposed research is to generate chemical tools that will expand our understanding of ADP-ribosylation in neuronal physiology. ADP-ribosylation was originally thought to be catalyzed by a single enzyme, ARTD1 (ADP-ribosyltransferases 1), but a family of 17 proteins is now recognized in humans that shares structural homology to the ARTD1 catalytic domain. ARTD1, and perhaps other ARTDs, play essential roles in cellular pathways in neurons that mediate long-term memory (LTM)~ however, their roles in these processes are not well understood. Moreover, the direct protein targets of individual ARTDs in neurons are not known, hindering our ability to fully delineate the pathway from ARTD activation to LTM. Our current lack of understanding of the specific role of ARTD1, and other ARTDs, in neurons and in other cell types has been severely limited by the lack of inhibitors of individual family members and the inability to identify the direct targets fr individual ARTDs in a cellular context. To overcome these limitations, this application describes, for the first time, the design and synthesis of (1) mono-selective inhibitors and (2) orthogonal NAD+ substrate analogs of ARTD1 mutants that are engineered to contain a unique pocket absent from wild-type ARTDs, but retain enzymatic activity. These orthogonal NAD+ analogs will be used for the identification of direct targets of ARTD1 in neurons. While initial studies wll focus on the role of ARTD1 in neurons, we anticipate that our strategy can be generalized to other ARTDs, thereby potentially providing unprecedented insights into their roles in physiology and pathophysiology.
Many physiologically important events are regulated at the level of posttranslational modification. ADP- ribosylation is increasingly being recognized as an essential posttranslational modification that plays an important role in neuronal physiology, where it regulates learning and memory, and pathophysiology, where its dysregulation leads to early onset neurodegeneration. Current strategies to study ADP-ribosylation are insufficient to provide a comprehensive view of its role in cellular processes tht underlying these events. This proposal describes, for the first time, the development of chemical tools that will yield new insights into the role of ADP-ribosylation in long-term memory formation, and will illuminate important molecular targets for therapeutic intervention in neurodegeneration.
|Thorsell, Ann-Gerd; Ekblad, Torun; Karlberg, Tobias et al. (2017) Structural Basis for Potency and Promiscuity in Poly(ADP-ribose) Polymerase (PARP) and Tankyrase Inhibitors. J Med Chem 60:1262-1271|
|Carter-O'Connell, Ian; Jin, Haihong; Morgan, Rory K et al. (2016) Identifying Family-Member-Specific Targets of Mono-ARTDs by Using a Chemical Genetics Approach. Cell Rep 14:621-631|
|Huang, Jeffrey Y; Wang, Kang; Vermehren-Schmaedick, Anke et al. (2016) PARP6 is a Regulator of Hippocampal Dendritic Morphogenesis. Sci Rep 6:18512|
|Cambronne, Xiaolu A; Stewart, Melissa L; Kim, DongHo et al. (2016) Biosensor reveals multiple sources for mitochondrial NAD?. Science 352:1474-7|
|Morgan, Rory K; Carter-O'Connell, Ian; Cohen, Michael S (2015) Selective inhibition of PARP10 using a chemical genetics strategy. Bioorg Med Chem Lett 25:4770-3|
|Carter-O'Connell, Ian; Cohen, Michael S (2015) Identifying Direct Protein Targets of Poly-ADP-Ribose Polymerases (PARPs) Using Engineered PARP Variants-Orthogonal Nicotinamide Adenine Dinucleotide (NAD+) Analog Pairs. Curr Protoc Chem Biol 7:121-39|
|Morgan, Rory K; Cohen, Michael S (2015) A Clickable Aminooxy Probe for Monitoring Cellular ADP-Ribosylation. ACS Chem Biol 10:1778-84|
|Carter-O'Connell, Ian; Jin, Haihong; Morgan, Rory K et al. (2014) Engineering the substrate specificity of ADP-ribosyltransferases for identifying direct protein targets. J Am Chem Soc 136:5201-4|