Caspases 8 and 10 Exosite Scan, Apoptosis-versus-proliferation Role Differentiation:

Friday, October 28, 2011
Hall 1-2 (San Jose Convention Center)
Roberto Nelson , University of California San Diego, San Diego, CA
Guy Salvesen, PhD , Biomedical Department, Sanford Burnham Medical Research Institute, La Jolla, CA
Interest in allosteric inhibition has been stimulated by recurring difficulties in the development of highly selective active-site protease inhibitors. This is in part due to overlap of active-site functionality as well as low active-site specificity. Defects in caspase-8 proteins have been shown to be directly related to the failure of the cellular apoptosis system. This project’s aim is to improve the understanding of specific mechanisms through which caspases 8 selectively interact with distinct proteins in order to allow for proper intervention in cellular apoptosis, with the purpose of fighting cancer. My project’s goal is to engineer protein-based substrates for the identification of specific exosites on caspase-8. Generation of allosteric substrates will be achieved by synthesis of protein domains via formation of intein hybrids attached to the C-terminus of the protein domain of interest.  I will determine catalytic parameters kcat and Km of the protein substrate reporters via spectroscopy by comparisons between conventional substrate velocity plots and optimal tetrapeptide substrates.