Peptidomic analysis of the larval Drosophila melanogaster central nervous system by two-dimensional capillary liquid chromatography quadrupole time-of-flight mass spectrometry.
Peptides are the largest class of signalling molecules found in animals. Nevertheless, in most proteomic studies peptides are overlooked since they literally fall through the mazes of the net. In analogy with proteomics technology, where all proteins expressed in a cell or tissue are analyzed, the peptidomic approach aims at the simultaneous visualization and identification of the whole peptidome of a cell or tissue, i.e. all expressed peptides with their post-translational modifications. In this paper we describe the analysis of the larval fruit fly central nervous system using two-dimensional capillary liquid chromatography/quadrupole time-of-flight tandem mass spectrometry (LC/Q-TOF-MS/MS. Using the central nervous systems of only 50 larval Drosophila as starting material, we identified 38 peptides in a single analysis, 20 of which were not detected in a previous study that reported on the one-dimensional capillary LC/MS/MS analysis of the same tissue. Among the 38 sequenced peptides, some originate from precursors, such as the tachykinin and the IFamide precursor that were entirely missed in the first study. This clearly demonstrates that the two-dimensional capillary LC approach enhances the coverage of the peptidomic analysis.
Baggerman
Geert
G
Laboratory of Developmental Physiology, Genomics and Proteomics, K.U. Leuven, Naamsestraat 59, B-3000 Leuven, Belgium. geert.baggerman@bio.kuleuven.ac.be
Boonen
Kurt
K
Verleyen
Peter
P
De Loof
Arnold
A
Schoofs
Liliane
L
D000595:Amino Acid Sequence; D000818:Animals; D001923:Brain Chemistry; D002490:Central Nervous System / Q000737:chemistry / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D004331:Drosophila melanogaster / Q000737:chemistry / Q000378:metabolism*; D019835:FMRFamide / Q000031:analogs & derivatives / Q000032:analysis; D007301:Insect Hormones / Q000032:analysis; D007814:Larva / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000032:analysis; D009842:Oligopeptides / Q000032:analysis; D010455:Peptides / Q000032:analysis; D011498:Protein Precursors / Q000032:analysis / Q000737:chemistry; D040901:Proteomics; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives* / Q000032:analysis
D007301:Insect Hormones; D009479:Neuropeptides; D009842:Oligopeptides; D010455:Peptides; D011498:Protein Precursors; C044833:adipokinetic hormone; C076523:allatostatin; C088629:leucokinin 1; D019835:FMRFamide; D011761:Pyrrolidonecarboxylic Acid