A molecular scanner to automate proteomic research and to display proteome images

Article Article scientifique Article A molecular scanner to automate proteomic research and to display proteome images eng 1999 author Binz Pierre-Alain 122663 author Muller Markus Johann 125264 author Walther Daniel 83450 author Bienvenut Willy-Vincent 503954 author Gras Robin 126613 author Hoogland Christine 503821 author Bouchet Gérard 123446 author Gasteiger Elisabeth 101626 author Fabbretti Robert 29445 author Gay Steven Daryl 28765 author Palagi Graf Patricia 119158 author Wilkins Marc 111772 author Rouge Véronique author Tonella Luisa 78859 author Paesano Salvo 133808 author Rossellat Gérald 133395 author Karmime Abderrahim 29133 author Bairoch Amos Marc 0000-0003-2826-6444 21964 author Sanchez Jean-Charles 0000-0002-8733-5932 24399 author Appel Ron David 22305 author Hochstrasser Denis 26422 4981-4988 10.1021/ac990449e 10565287 0003-2700 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 still 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 annoated 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. Automation Blood Proteins/analysis/chemistry Electrophoresis, Gel, Two-Dimensional/methods Electrophoresis, Polyacrylamide Gel/methods Humans Image Processing, Computer-Assisted/instrumentation/methods Peptide Mapping/methods Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods Trypsin Inhibitor, Kunitz Soybean/analysis/chemistry Dewey 576 136 Département de biologie structurale et bioinformatique 21E false 57 Calipho 80 211 Auto-immunité, inflammation et athérosclérose 270 369 Biomarqueurs 635 Analytical chemistry 71 21 234c1e74-060b-4c01-92e0-987230eab20e Article (Published version) Public pub_094_10565287.pdf 318300 application/pdf openAccess Faculté de médecine / Section de médecine fondamentale / Département de biologie structurale et bioinformatique

Title	Published in	Access level	OA Policy	Year	Views	Downloads

A molecular scanner to automate proteomic research and to display proteome images	Analytical chemistry			1999	646	866

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Rossellat, Gérald