학술논문
A Molecular Scanner To Automate Proteomic Research and To Display Proteome Images
Document Type
Academic Journal
Author
Binz, Pierre-Alain; Muller, Markus; Walther, Daniel; Bienvenut, Willy V.; Gras, Robin; Hoogland, Christine; Bouchet, Gerard; Gasteiger, Elisabeth; Fabbretti, Roberto; Gay, Steven; Palagi, Patricia; Wilkins, Marc R.; Rouge, Veronique; Tonella, Luisa; Paesano, Salvo; Rossellat, Gerald; Karmime, Abderrahim; Bairoch, Amos; Sanchez, Jean-Charles; Appel, Ron D.; Hochstrasser, Denis F.
Source
Analytical Chemistry. Nov 1, 1999, Vol. 71 Issue 21, p4981, 8 p.
Subject
Language
ISSN
0003-2700
Abstract
Identification and characterization of all proteins expressed by a genome in biological samples represent major challenges in proteomics. Today's commonly used high-throughput approaches combine two-dimensional electrophoresis (2-DE) with peptide mass fingerprinting (PMF) analysis. Although automation is often possible, a number of limitations stir adversely affect the rate of protein identification and annotation in 2-DE databases: the sequential excision process of pieces of gel containing protein; the enzymatic digestion step; the interpretation of mass spectra (reliability of identifications); and the manual updating of 2-DE databases. We present a highly automated method that generates a fully annotated 2-DE map. Using a parallel process, all proteins of a 2-DE are first simultaneously digested proteolytically and electro-transferred onto a poly(vinylidene difluoride) membrane. The membrane is then directly scanned by MALDI-TOF MS. After automated protein identification from the obtained peptide mass fingerprints using PeptIdent software (http://www. expasy, ch/tools/peptident, html), a fully annotated 2-D map is created on-line. It is a multidimensional representation of a proteome that contains interpreted PMF data in addition to protein identification results. This 'MS-imaging' method represents a major step toward the development of a clinical molecular scanner.