MALDI/MS peptide mass fingerprinting for proteome analysis: identification of hydrophobic proteins attached to eucaryote keratinocyte cytoplasmic membrane using different matrices in concert

Affordable Access

MALDI/MS peptide mass fingerprinting for proteome analysis: identification of hydrophobic proteins attached to eucaryote keratinocyte cytoplasmic membrane using different matrices in concert

Publisher
BioMed Central
Publication Date
May 06, 2003
Source
PMC
Keywords
Disciplines
  • Biology
License
Unknown

Abstract

1477-5956-1-2.fm ral ss BioMed CentProteome Science Open AcceResearch MALDI/MS peptide mass fingerprinting for proteome analysis: identification of hydrophobic proteins attached to eucaryote keratinocyte cytoplasmic membrane using different matrices in concert Florence Gonnet*1, Gilles Lemaître2,3, Gilles Waksman2,3 and Jeanine Tortajada1 Address: 1Laboratoire Analyse et Environnement, UMR CNRS 8587, Université d'Evry-Val-d'Essonne, Bd. F. Mitterrand, 91025 Evry Cedex, France, 2Université d'Evry-val d'Essonne, EA 2541, 2 rue Gaston Crémieux, 91057 Evry Cedex, France and 3Service de Génomique Fonctionnelle, CEA, 2 rue Gaston Crémieux, 91057 Evry Cedex, France Email: Florence Gonnet* - [email protected]; Gilles Lemaître - [email protected]; Gilles Waksman - [email protected]; Jeanine Tortajada - [email protected] * Corresponding author Abstract Background: MALDI-TOF-MS has become an important analytical tool in the identification of proteins and evaluation of their role in biological processes. A typical protocol consists of sample purification, separation of proteins by 2D-PAGE, enzymatic digestion and identification of proteins by peptide mass fingerprint. Unfortunately, this approach is not appropriate for the identification of membrane or low or high pI proteins. An alternative technique uses 1D-PAGE, which results in a mixture of proteins in each gel band. The direct analysis of the proteolytic digestion of this mixture is often problematic because of poor peptide detection and consequent poor sequence coverage in databases. Sequence coverage can be improved through the combination of several matrices. Results: The aim of this study was to trust the MALDI analysis of complex biological samples, in order to identify proteins that interact with the membrane network of keratinocytes. Peptides obtained from protein trypsin digestions may have either hydrophobic or hydrophilic sections, in which case, the direct analys

Report this publication

Statistics

Seen <100 times