Signal Traitment and Virtual Images Production (1/2) View Full Text


Ontology type: schema:Chapter     


Chapter Info

DATE

2005

AUTHORS

PA. Binz , M. Muller , D. Walther , WV. Bienvenut , R. Gras , C. Hoogland , G. Bouchet , E. Gasteiger , R. Fabbretti , S. Gay , P. Palagi , MR. Wilkins , V. Rouge , L. Tonella , S. Paesano , G. Rossellat , A. Karmime , A. Bairoch , JC. Sanchez , RD. Appel , D F Hochstrasser

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 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 annotated 2-DE map. Using a parallel process, all proteins of a 2-DE are first simultaneously digested proteolytically and electro-transferred onto a polyvinylidene difluoride (PVDF) 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 online. It is a multi-dimensional representation of a proteome, that contains interpreted PMF data in addition to protein identification results. This “MS-imaging” method represents a major step towards the development of a clinical molecular scanner. More... »

PAGES

151-168

References to SciGraph publications

Book

TITLE

Acceleration and Improvement of Protein Identification by Mass Spectrometry

ISBN

1-4020-3318-4

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/1-4020-3319-2_4

DOI

http://dx.doi.org/10.1007/1-4020-3319-2_4

DIMENSIONS

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