MASS-SPECTROMETRIC IDENTIFICATION AND MICROCHARACTERIZATION OF PROTEINS FROM ELECTROPHORETIC GELS - STRATEGIES AND APPLICATIONS

The entire genomic DNA sequences of a number of prokaryotic and eukary otic species are now available and many more, including the human geno me, will be completed in the near future. The state-of-life of a cell at any given time, however, is defined by its protein composition, i.e ., its proteome. Gel electrophoresis, mass spectrometry, and bioinform atics will be important tools for protein and proteome analysis in the post-genome era. Protein identification from electrophoretic gels by mass spectrometric peptide mapping or peptide sequencing combined with sequence database searching is established and has been applied to nu merous biological systems. We describe current strategies and selected applications in molecular and cell biology. The next challenges are d etailed structure/function analyses, which include studying the molecu lar composition of multiprotein complexes and characterization of seco ndary modifications of proteins. The advantages and limitations of a n umber of mass spectrometry-based strategies designed for microcharacte rization of low amounts of protein from electrophoretic gels are discu ssed and illustrated by examples. Proteins Suppl. 2:74-89, 1998, (C) 1 998 Wiley-Liss, Inc.