title: Evaluation of combined matrix-assisted laser desorption/ionization time-of-flight and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry experiments for peptide mass fingerprinting analysis.
abstract: Peptide Mass Fingerprinting (PMF) is still of significant interest in proteomics because it allows a large number of complex samples to be rapidly screened and characterized. The main part of post-translational modifications is generally preserved. In some specific cases, PMF suffers from ambiguous or unsuccessful identification. In order to improve its reliability, a combined approach using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and matrix-assisted laser desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICRMS) was evaluated. The study was carried out on bovine serum albumin (BSA) digest. The influence of several important parameters (the matrix, the sample preparation method, the amount of the analyte) on the MOWSE score and the protein sequence coverage were evaluated to allow the identification of specific effects. A careful investigation of the sequence coverage obtained by each kind of experiment ensured the detection of specific peptides for each experimental condition. Results highlighted that DHB-FTICRMS and DHB- or CHCA-TOFMS are the most suited combinations of experimental conditions to achieve PMF analysis. The association (convolution) of the data obtained by each of these techniques ensured a significant increase in the MOWSE score and the protein sequence coverage.
authors.affiliation: Laboratoire de Spectrométrie de Masse et de Chimie Laser (LSMCL), Institut Jean Barriol - Fédération de Recherche 2843, Université Paul Verlaine - Metz, 1, Boulevard Arago, F-57078 Metz technopole Cedex 03, France.
authors.forename: David
authors.initials: D
authors.lastname: da Silva
authors.forename: Thierry
authors.initials: T
authors.lastname: Wasselin
authors.forename: Vincent
authors.initials: V
authors.lastname: Carré
authors.forename: Patrick
authors.initials: P
authors.lastname: Chaimbault
authors.forename: Lina
authors.initials: L
authors.lastname: Bezdetnaya
authors.forename: Benoît
authors.initials: B
authors.lastname: Maunit
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Muller
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D003373:Coumaric Acids / Q000737:chemistry; D005583:Fourier Analysis; D005841:Gentisates / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010449:Peptide Mapping / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
chemical_list: D003373:Coumaric Acids; D005841:Gentisates; D012710:Serum Albumin, Bovine; C007175:alpha-cyano-4-hydroxycinnamate; C010925:2,5-dihydroxybenzoic acid