- Day 2, May 16（Thu.） 09:50-10:10 Room E (201)
Development of Novel Methods to Enrich Protein C-terminal Peptides by Metal Oxide Chromatography
Since the protein expression profiles between organs, tissues, cell types, or diseases often depend on their proteoforms such as proteins with different termini, it is important to comprehensively obtain information on protein termini in order to understand the protein functions. In shotgun proteomics, however, it is not so easy to measure the digested peptides containing protein N or C-termini in the presence of huge amounts of other peptides. In this study, we developed novel methods to enrich protein C-terminal peptides in order to profile the protein terminomes. We used V8 protease coupled with metal oxide chromatography using titania or cerium oxide to exclude the internal peptides. Compared to the previous methods based on series of chemical reactions, this one-step method is expected to be simple, sensitive and reproducible. We optimized some parameters such as metal and pH during the enrichment process, suggesting that each metal has different optimum pH.