Summary Technology developed at OHSU provides a novel method to identify proteins regulated by PARP mono-ADP-ribosylation (MARylation), a post-translational modification carried out by the majority of enzymes within this family. This tool has the ability to link such PARPs to specific targets and broader biological pathways, thereby increasing our understanding of their signaling cascades to improve outcomes of translational research in support of drug development. Technology Overview PARPs are powerful pharmaceutical targets due to their diverse physiological functions, but linking the large number of PARP family members to their specific protein targets and pathways is a significant challenge. Dr. Cohen’s laboratory has developed a novel platform to efficiently identify protein targets of individual PARPs that catalyze MARylation, so called monoPARPs. MARylation regulates numerous cellular processes and consists of the transfer of ADP-ribose from an NAD molecule to the targeted protein. The current technology is based on genetically engineering monoPARP binding away from endogenous NAD, to a tagged NAD analog not used by wild-type PARPs (see Figure). Individual monoPARPs can then be genetically modified in a cell line of interest and incubated with the modified NAD, after which cell lysates are collected for tag retrieval and mass spectrometry identification of targeted proteins. Investigation of 4 monoPARPs in HEK293 cells identified between 14-500 unique protein targets for each monoPARP studied. This novel platform can be used to link PARP biology to specific monoPARPs, increasing the potential to design more selective and specific PARP regulatory molecules for translational research and pharmaceutical development in fields such as neurodegeneration and cancer.
Publications Carter-O’Connell et al., “Engineering the substrate specificity of ADP-Ribosyltransferases for Identifying Direct Protein Targets.” Journal of the American Chemical Society 136(2014): 5201-5204. Link Carter-O’Connell et al., “Identifying Family-Member-Specific Targets of Mono-ARTDs by Using a Chemical Genetics Approach.” Cell Reports 14(2016): 621-631. Link Licensing Opportunity This technology is available for licensing.