Aisosa Omere / Chemistry & Biochemistry / Faculty Mentor: Joseph Buonomo
Contemporary single-molecule protein sequencing relies on tandem mass spectrometry (LC-MS/MS), employing either top-down or bottom-up proteomics. Bottom-up approaches, which involve proteolytic fragmentation, can lose both post-translational modification data and the contextual integrity of the proteoform. While top-down methods analyze intact proteins, each approach is limited by high costs, low throughput, and large sample requirements, which hinder the analysis of low-abundance proteoforms. To address these challenges, we propose a novel Edman-based multiplexing protein sequencing strategy. This suite of technologies utilizes chemoselective probes and advanced computational techniques to enable comprehensive and quantitative analyses of proteoforms. This presentation will focus on the foundational aspect of our technology: the development of a C-terminal-specific ligation strategy, a critical step that enables subsequent Edman degradation and precise protein sequencing. Contemporary immobilization strategies utilize unnatural amino acids to facilitate the adherence of peptides to solid surfaces; otherwise, the results yield very low returns. Here, we report our successful ability to immobilize a native peptide onto a solid surface.
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