A modified hybrid Qq time of flight mass spectrometer for analysis of multiprotein complexes

Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.

We propose to extend significantly our mass spectrometry investigations of non-covalent complexes taking the mass spectrometer into new areas of science by allowing us to access complexes present in lower amounts and to induce more extensive fragmentation. Together these attributes will allow us to: observe directly intact complexes that have been predicted to exist from interaction networks; allow nearest neighbour deductions within the complex; distinguish transient interactions due to temporal changes in the cell reaction cycle; allow definition of metal/nucleic acid binding associated with the complexes not identified by existing methods; identify low levels of proteins that co purify with complexes; when coupled with cross-linking approaches will allow distant constraints to enable us to provide detailed structural models; will also allow us to identify post translational modifications; to define complexes involved in various stages of splicing reactions; probe interactions in both pro- and eukaryotic ribosomes in different functional states; characterise complexes involved in protein degradation in the proteasome from both pro and eukaryotic sources. Specifically we will focus on applying these approaches to complexes involved in translation, RNA splicing and protein degradation although we anticipate a wide network of collaborations with other researchers nationally and internationally.