Biophysical analysis of macromolecular associations. (360G-Wellcome-082897_Z_07_Z)

This is an application to replace defunct equipment at York with new instrumentation that will service the needs of the 4 Trust-funded applicants and a further 14 staff in the Departments of Biology and Chemistry in their investigations of biomedically and biologically important proteins. Specifically, we request a surface plasmon resonance instrument (SPR) and electrospray ionisation mass spectrometer (ESI-MS), which will together be used to deconstruct the associations of a wide variety of pr otein complexes and so provide much needed biophysical information to complement ongoing structural work. The key objectives of the proposal are: 1. To use SPR to investigate the assembly and binding affinities of protein complexes and probe their underlying kinetics. 2. To use native state ESI-MS to measure molecular weights of complexes from which we can infer subunit and ligand stoichiometries and oligomeric structure. These approaches will allow us to address fundamental questions in ongoing, Trust-funded work that have hitherto been impossible at York. The areas of research that will benefit from the new equipment include host-parasite interactions, host-bacterial pathogen interactions, viral DNA packaging, antibacterial protein translocation, bacterial transcription and cell division, mammalian DNA replication, archaeal DNA segregation, bone cell signalling and protein sorting.