Incorporating Biological Mass Spectrometry into Undergraduate Teaching Labs, Part 3: De Novo Peptide Sequencing Using Electrospray Tandem Mass Spectrometry

摘要

Tandem mass spectrometry (MSMS) is an important techniqueroutinely used to provide structural information about analytes of interest (1). While undergraduate curricula often include a discussion of mass spectrometry, tandem mass spectrometry is often not taught. Even fewer institutions incorporate MSMS experiments into their lab curriculum. A search of previous lab experiments utilizing tandem mass spectrometry in this Journal identified only four examples (2–5). One of these uses GC–MS to study chlorobenzenes (2). Two other experiments utilize LC–MS techniques to analyze small molecules. One of these investigates gas-phase basicities of individual amino acids (3) and the other focuses on environmental contaminants found in fish liver (4). While small molecule analysis is an important application of tandem mass spectrometry, so too is using MSMS techniques to study proteins and peptides. The final manuscript describes using tandem mass spectrometry to analyze tryptic peptides resulting from a protein digestion (5). The tandem mass spectrometry data are not analyzed de novo, but rather are compared to the expected fragmentation pattern for the peptide being analyzed as a confirmation of protein identity.