抄録
We describe here an integrated protein identification system constructed by automated two-dimensional liquid chromatography and electrospray ionization-tandem mass spectrometry on a hybrid mass spectrometer (2DLC-MS/MS), coupled with a data process-analysis system to identify proteins on a genome database. Application of this system to the analysis of the proteome of E. coli and a multi-cellular model organism C. elegans allowed identification of 1,500-2,300 proteins including those having extreme pI (<4, >10) or large Mr (>200 kDa). The identified proteins also included many membrane proteins such as cell surface receptor and cell adhesion molecules. Retrieval of the large volume of MS/MS data further elucidated a variety of post-translational modifications in proteins, such as N-terminal acetylation and a site-specific phosphorylation of Ser, Thr or Tyr. The 2DLC-MS/MS system could also be applied to the comprehensive, large-scale analysis of glycoproteins after specific isotope tagging of glycopeptides captured by lectin-affinity chromatography.
