Bio/Technology
14, 61 - 65 (1996)
doi:10.1038/nbt0196-61
From Proteins to Proteomes: Large Scale Protein Identification by Two-Dimensional Electrophoresis and Arnino Acid AnalysisMarc R. Wilkins1, Christian Pasquali2, Ron D. Appel2, Keli Ou1, Olivier Golaz2, Jean-Charles Sanchez2, Jun X. Yan1, Andrew. A. Gooley1, Graham Hughes2, Ian Humphery-Smith3, Keith L. Williams1, *
& Denis F. Hochstrasser2
1Macquarie University Centre for Analytical Biotechnology, Macquarie University, Sydney, NSW 2109, Australia.
2Central Clinical Chemistry Laboratory and Medical Computing Centre of the University of Geneva, CH 1211 Geneva 14, Switzerland.
3Department of Microbiology, University of Sydney, NSW, 2006, Australia.
*e-mail: keith.williams@mq.edu.au Separation and identification of proteins by two-dimensional (2-D) electrophoresis can be used for protein-based gene expression analysis. In this report single protein spots, from polyvinylidene difluoride blots of micropreparative E. coli 2-D gels, were rapidly and economically identified by matching their amino acid composition, estimated pI and molecular weight against all E. coli entries in the SWISS-PROT database. Thirty proteins from an E. coli 2-D map were analyzed and identities assigned. Three of the proteins were unknown. By protein sequencing analysis, 20 of the 27 proteins were correctly identified. Importantly, correct identifications showed unambiguous "correct" score patterns. While incorrect protein identifications also showed distinctive score patterns, indicating that protein must be identified by other means. These techniques allow large-scale screening of the protein complement of simple organisms, or tissues in normal and disease states. The computer program described here is accessible via the World Wide Web at URL address (http://expasy.hcuge.ch/).
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