title: Seed storage proteins as a system for teaching protein identification by mass spectrometry in biochemistry laboratory.
abstract: Mass spectrometry (MS) has become an important tool in studying biological systems. One application is the identification of proteins and peptides by the matching of peptide and peptide fragment masses to the sequences of proteins in protein sequence databases. Often prior protein separation of complex protein mixtures by 2D-PAGE is needed, requiring more time and expertise than instructors of large laboratory classes can devote. We have developed an experimental module for our Biochemistry Laboratory course that engages students in MS-based protein identification following protein separation by one-dimensional SDS-PAGE, a technique that is usually taught in this type of course. The module is based on soybean seed storage proteins, a relatively simple mixture of proteins present in high levels in the seed, allowing the identification of the main protein bands by MS/MS and in some cases, even by peptide mass fingerprinting. Students can identify their protein bands using software available on the Internet, and are challenged to deduce post-translational modifications that have occurred upon germination. A collection of mass spectral data and tutorials that can be used as a stand-alone computer-based laboratory module were also assembled.
authors.affiliation: Department of Biological Sciences, State University of New York at Binghamton, Binghamton, New York, 13902-6000, USA. kwilson@binghamton.edu
authors.forename: Karl A
authors.initials: KA
authors.lastname: Wilson
authors.forename: Anna
authors.initials: A
authors.lastname: Tan-Wilson
mesh_terms: D000595:Amino Acid Sequence; D001671:Biochemistry / Q000193:education*; D004591:Electrophoresis, Polyacrylamide Gel; D007753:Laboratories; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D055314:Seed Storage Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D013025:Glycine max; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013663:Teaching; D014357:Trypsin / Q000378:metabolism
chemical_list: D010455:Peptides; D055314:Seed Storage Proteins; D014357:Trypsin