Identifying SAM-binding proteins in D.melanogaster

The aim of the project is to identify proteins the ustilize S-adenosyl methionine as cofactor for performing methylation of target molecules in D. melanogaster S2 cells. For this purpose, we applied a small-molecule capturing protocol using a SAH-capture compound with UV-activated crosslinker and biotin immobilization groups. We tested the protein affinity with and without crosslinking for two different photo-activatable crosslinkers benzophenone and nitrobenzene, respectively, as well as different concentrations of the capture compound. As control we incubated the cell lysate with the streptavidin beads without adding the capture compound. Two specific capture compounds containing a S-adenosyl homocysteine as capturing functionality and either benzophenone or azo-benzene as reactive crosslinkers as well as a biotin group for affinity purification were tested in this protocol. One sample representing a HeLa cell lysate (Waters) under equal conditions was added to the dataset.