Comparison of electrospray and matrix-assisted laser desorption ionization on the same hybrid quadrupole time-of-flight tandem mass spectrometer: application to bidimensional liquid chromatography of proteins from bovine milk fraction.
Recently, two ionization sources, electrospray (ESI) and matrix-assisted laser desorption (MALDI) have been used in parallel to exploit their complementary nature and to increase proteome coverage. In this study, a method using bidimensional (2D) nanoLC coupled online with ESI quadrupole time-of-flight (Q-TOF) with the simultaneous collection of fractions for analyses by LC-MALDI Q-TOF-MS/MS was developed. A total of 39 bovine proteins were identified to a high degree of confidence. To help in differentiating peptide detection following ESI and MALDI with the same mass spectrometer, we compared physico-chemical characteristics of the peptides (molecular mass, charge and size) by principal component analysis (PCA) and analysis of variance on the results of PCA. More hydrophobic peptides with a wider mass coverage were identified when ESI was used, whereas more basic and smaller peptides were identified when MALDI was used. However, the generally accepted differentiation between ESI and MALDI according to the presence of basic amino acids residues Lys and Arg and the ratio Lys/Arg was not shown as significant in this study. Moreover, we pointed out the importance of the type of mass spectrometer used in complement to both ionization sources for achieving a global increase of proteome coverage.
Mollé
Daniel
D
INRA, UMR1253, Rennes, France.
Jardin
Julien
J
Piot
Michel
M
Pasco
Maryvonne
M
Léonil
Joëlle
J
Gagnaire
Valérie
V
D000704:Analysis of Variance; D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002853:Chromatography, Liquid / Q000379:methods*; D004867:Equipment Design; D057927:Hydrophobic and Hydrophilic Interactions; D013058:Mass Spectrometry / Q000379:methods*; D008894:Milk Proteins / Q000737:chemistry / Q000302:isolation & purification*; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D025341:Principal Component Analysis
D008894:Milk Proteins; D010455:Peptides