Activity Based Protein Profiling
Activity based probes, which mimic an enzyme substrate or protein binding partner but contain a chemical trap, are powerful tools in biological research. Such probes have been employed widely for both basic research and guiding drug discovery efforts. They are used to discover and characterise novel enzymatic activities, inhibitors, biomarkers and cellular localisation.
The advantages of a “chemical proteomics” approach using activity-based probes over biochemical assays lie in the fact that they allow the selective study of enzymes in their native environment. Alterations with cell type and disease state and have a huge impact on protein function and selectivity, an activity based profiling approach will be sensitive to these factors while still maintaining a highly specific readout, affording the best of both worlds between biochemical and phenotypic assays.
Research in the laboratory focuses on the synthesis of new activity based probes for chemical biology and molecular physiology applications. Current research focuses on DNA damage repair and deubiquitinating enzymes (DUBs) as two key regulatory classes of enzyme.
There is growing evidence for a pivotal role of DUBs in pathologies including infection, autoimmunity, cancer and neurodegeneration. DNA damage repair enzymes play crucial roles in cell maintenance and response to cancer treatments.
Techniques to make the probes range from protein expression and modification to right through to purely synthetic approaches. An activity based proteomics approach allows us to study these key enzymes in their native environment, examining how activity levels change with cellular state.
Relevant Publications
McKenna, S. M.; Fay, E. M.; McGouran, J. F. Flipping the switch: Innovations in inducible probes for protein profiling ACS Chem. Biol, 2021
Taylor, N. C.; McGouran, J. F. Investigating eosin Y as a photcatalyst for the radical-dependent activity-based probing of deubiquitinating enzymes, Organic & Biomolecular Chemistry, 2021
Taylor, N. C.; Hessman, G; Kramer, H; McGouran, J. F. Probing enzyme activity-a radical approach, Chemical Science, 2020
Taylor, N. C.; McGouran, J. F. Strategies to target specific components of the ubiquitin conjugation/deconjugation machinery, Frontiers in Chemistry, 2019,
Turnbull AP, Ioannidis S, Krajewski WW, PintoFernandez A, Heride C, Martin ACL, Tonkin LM, Townsend EC, Buker SM, Lancia Jr DR, Caravella JA, Toms AV, Charlton TM, Lahdenranta J, Wilker E, Follows BC, Evans NJ, Stead L, Alli C, Zarayskiy VV, Talbot AC, Buckmelter AJ, Wang M, McKinnon CL, Saab F, McGouran JF, Century H, Gersch M, Pittman MS, Marshall CG, Raynham TM, Simcox M, Stewart LMD, McLoughlin SB, Escobedo JA, Bair KW, Dinsmore CJ, Hammonds TR, Kim S, Urbé S, Clague MJ, Kessler BM, Komander D, Molecular basis of USP7 inhibition by selective small molecule inhibitors, Nature 2017,
McGouran JF, Gaertner SR, Altun M, Kramer HB, Kessler BM, Deubiquitinating enzyme specificity for ubiquitin chain topology profiled by di-ubiquitin activity probes., Chemistry & biology, 20, (12), 2013, 1447-55
Lercher L, McGouran JF, Kessler BM, Schofield CJ, Davis BG, DNA modification under mild conditions by Suzuki-Miyaura cross-coupling for the generation of functional probes., Angewandte Chemie (International ed. in English), 52, (40), 2013, 10553-8
Bush J, Walport L, McGouran J, Leung I, Berridge G, Van Berkel S, Basak A, Kessler B, Schofield C. The Ugi four-component reaction enables expedient synthesis and comparison of photoaffinity probes, Chemical Science, 4, (11), 2013, 4115-4120
McGouran JF, Kramer HB, Mackeen MM, di Gleria K, Altun M, Kessler BM, Fluorescence-based active site probes for profiling deubiquitinating enzymes., Organic & biomolecular chemistry, 10, (17), 2012, 3379-83
Altun M, Kramer HB, Willems LI, McDermott JL, Leach CA, Goldenberg SJ, Kumar KG, Konietzny R, Fischer R, Kogan E, Mackeen MM, McGouran J, Khoronenkova SV, Parsons JL, Dianov GL, Nicholson B, Kessler BM, Activity-based chemical proteomics accelerates inhibitor development for deubiquitylating enzymes., Chemistry & biology, 18, (11), 2011, 1401-12