My role within the group has been to develop new tools and methods for cross-linking/mass spectrometry (CLMS). My focus is to develop ways of increasing both data-density, in areas where data can already be obtained, as well as obtaining data where current CLMS methods are limited.
One development has been the use of UV-photoactivatable heterobifunctional cross-linkers to increase data density several-fold. Resulting high-density CLMS data can be used in combination with computational biology, to generate highly accurate models (2.5 Å RMSD to X-ray crystal structure), of protein structure analysed in the context of human blood serum. Development of a complimentary benzophenone CLMS photochemistry shows that different photoactivatable groups display orthogonal directionality.
Where CLMS data is currently missing/unobservable, I am looking at ways to access it. We are developing photo-AA technology for insights into protein structure from living cells. Coming from a synthetic peptide chemistry background, I am looking at ways to exploit chemical methods to provide us with new functionalised cross-linkers as well as a variety of functionalised solid-supports, that will provide us with a complete peptide enrichment system.
- Belsom A, Mudd G, Giese S, Auer M, Rappsilber J. Complimentary benzophenone cross-linking/mass spectrometry photochemistry. Anal Chem. 2017 May 16;89(10):5319-5324. doi:10.1021/acs.analchem.6b04938
- Belsom A, Schneider M, Brock O, Rappsilber J. Blind evaluation of hybrid protein structure analysis methods based on cross-linking. Trends Biochem Sci. 2016 Jul;41(7):564-7. doi:10.1016/j.tibs.2016.05.005.
- Belsom A, Schneider M, Fischer L, Brock O, Rappsilber J. Serum albumin domain structures in human blood serum by mass spectrometry and computational biology. Mol Cell Proteomics. 2016 Mar;15(3):1105-16. doi: 10.1074/mcp.M115.048504.