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.

[1]  D. Nässel.  Tachykinin-related peptides in invertebrates: a review , 1999, Peptides.

[2]  S. Gygi,et al.  Quantitative analysis of complex protein mixtures using isotope-coded affinity tags , 1999, Nature Biotechnology.

[3]  R. Murali,et al.  Therapeutic peptides and peptidomimetics. , 1997, Current opinion in biotechnology.

[4]  W. Haddon,et al.  Leucosulfakinin, a sulfated insect neuropeptide with homology to gastrin and cholecystokinin. , 1986, Science.

[5]  J. Veenstra.  Isolation and structure of two gastrin/CCK-like neuropeptides from the American cockroach homologous to the leucosulfakinins , 1989, Neuropeptides.

[6]  L. Schoofs,et al.  Isolation, identification and synthesis of PDVDHFLRFamide (SchistoFLRFamide) in Locusta migratoria and its association with the male accessory glands, the salivary glands, the heart, and the oviduct , 1993, Peptides.

[7]  T. Lee,et al.  Amino acid sequence of CAP2b, an insect cardioacceleratory peptide from the tobacco hawkmoth Manduca sexta , 1995, FEBS letters.

[8]  P. Verhaert,et al.  Insect neuropeptide F (NPF)-related peptides: isolation from Colorado potato beetle (Leptinotarsa decemlineata) brain. , 1996, Insect biochemistry and molecular biology.

[9]  H. Dircksen,et al.  Ecdysis of decapod crustaceans is associated with a dramatic release of crustacean cardioactive peptide into the haemolymph. , 2000, The Journal of experimental biology.

[10]  T. K. Hayes,et al.  Isolation, identification and synthesis of locustamyoinhibiting peptide (LOM-MIP), a novel biologically active neuropeptide from Locusta migratoria , 1991, Regulatory Peptides.

[11]  L. Schoofs,et al.  Sulfakinins reduce food intake in the desert locust, Schistocerca gregaria. , 2000, Journal of insect physiology.

[12]  T. K. Hayes,et al.  Isolation, primary structure, and synthesis of locustapyrokinin: a myotropic peptide of Locusta migratoria. , 1991, General and comparative endocrinology.

[13]  R. Nichols,et al.  Spatial and temporal expression identify dromyosuppressin as a brain‐gut peptide in Drosophila melanogaster , 1993, The Journal of comparative neurology.

[14]  S. Tobe,et al.  Comparison of the allatostatin neuropeptide precursors in the distantly related cockroaches Periplaneta americana and Diploptera punctata. , 1995, European journal of biochemistry.

[15]  D. Kojima,et al.  Functional Interaction between Bovine Rhodopsin and G Protein Transducin* , 2002, The Journal of Biological Chemistry.

[16]  S. Tobe,et al.  Primary structure of four allatostatins: neuropeptide inhibitors of juvenile hormone synthesis. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[17]  K. Yagi,et al.  Callatostatins: neuropeptides from the blowfly Calliphora vomitoria with sequence homology to cockroach allatostatins. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Liliane Schoofs,et al.  Peptidomics in Drosophila melanogaster. , 2003, Briefings in functional genomics & proteomics.

[19]  D. Nässel,et al.  Neuronal expression of tachykinin‐related peptides and gene transcript during postembryonic development of Drosophila , 2003, The Journal of comparative neurology.

[20]  X. Bertagna,et al.  Proopiomelanocortin, a polypeptide precursor with multiple functions: from physiology to pathological conditions. , 2003, European journal of endocrinology.

[21]  M. Williamson,et al.  Molecular cloning and genomic organization of an allatostatin preprohormone from Drosophila melanogaster. , 2000, Biochemical and biophysical research communications.

[22]  R. Kellner,et al.  Identification of novel periviscerokinins from single neurohaemal release sites in insects MS/MS fragmentation complemented by Edman degradation. , 2000, European journal of biochemistry.

[23]  J. Veenstra.  Isolation and structure of the Drosophila corazonin gene. , 1994, Biochemical and biophysical research communications.

[24]  T. K. Hayes,et al.  Isolation and Identification of a Sulfakinin-Like Peptide, with Sequence Homology to Vertebrate Gastrin and Cholecystokinin, from the Brain of Locusta Migratoria , 1990 .

[25]  W. Haddon,et al.  Leucosulfakinin-II, a blocked sulfated insect neuropeptide with homology to cholecystokinin and gastrin. , 1986, Biochemical and biophysical research communications.

[26]  J. Broeck,et al.  Peptides in the Locusts, Locusta migratoria and Schistocerca gregaria 1 1 Taken in part from a paper presented at a satellite symposium on Insect Neuropeptides during the Seventh Annual Neuropeptide Conference, February 1–6, 1996, Breckenridge, CO. , 1997, Peptides.

[27]  D. Hunt,et al.  Isolation of NEB-LFamide, a novel myotropic neuropeptide from the grey fleshfly , 1996, Molecular and Cellular Endocrinology.

[28]  L. Schoofs,et al.  Pyrokinin neuropeptides in a crustacean. Isolation and identification in the white shrimp Penaeus vannamei. , 2001, European journal of biochemistry.

[29]  J. Rehfeld,et al.  The Sulfakinins of the Blowfly Calliphora Vomitoria , 1995 .

[30]  M. Williamson,et al.  Molecular cloning, genomic organization, and expression of a C-type (Manduca sexta-type) allatostatin preprohormone from Drosophila melanogaster. , 2001, Biochemical and biophysical research communications.

[31]  P. Taghert,et al.  Neuropeptides and neuropeptide receptors in the Drosophila melanogaster genome. , 2001, Genome research.

[32]  K. Gevaert,et al.  Exploring proteomes and analyzing protein processing by mass spectrometric identification of sorted N-terminal peptides , 2003, Nature Biotechnology.

[33]  J. Dow,et al.  Neuropeptide stimulation of the nitric oxide signaling pathway in Drosophila melanogaster Malpighian tubules. , 1997, The American journal of physiology.

[34]  P. Proost,et al.  Isolation and primary structure of two sulfakinin-like peptides from the fleshfly, Neobellieria bullata. , 1992, Comparative biochemistry and physiology. C, Comparative pharmacology and toxicology.

[35]  T. K. Hayes,et al.  Leucokinins, a new family of ion transport stimulators and inhibitors in insect Malpighian tubules. , 1989, Life sciences.

[36]  L. Schoofs,et al.  Identification of the gregarization-associated dark-pigmentotropin in locusts through an albino mutant. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[37]  Liliane Schoofs,et al.  SIFamide is a highly conserved neuropeptide: a comparative study in different insect species. , 2004, Biochemical and biophysical research communications.

[38]  T. K. Hayes,et al.  Locustatachykinin I and II, two novel insect neuropeptides with homology to peptides of the vertebrate tachykinin family , 1990, FEBS letters.

[39]  D. Garland,et al.  An immunocytochemical study of the FMRFamide neuropeptide gene products in Drosophila , 1993, The Journal of comparative neurology.

[40]  B. Devreese,et al.  Isolation and characterization of schistostatin-211–18 from the desert locust, Schistocerca gregaria: A truncated analog of schistostatin-2 , 1996, Regulatory Peptides.

[41]  J. Huybrechts,et al.  Diapausing Colorado potato beetles are devoid of short neuropeptide F I and II. , 2004, Biochemical and biophysical research communications.

[42]  H. Penzlin,et al.  Periviscerokinin (Pea-PVK): A novel myotropic neuropeptide from the perisympathetic organs of the American cockroach , 1995, Peptides.

[43]  S. Tobe,et al.  Two new allatostatins from the brains of Diploptera punctata. , 1994, Insect biochemistry and molecular biology.

[44]  J. Veenstra,et al.  Mono- and dibasic proteolytic cleavage sites in insect neuroendocrine peptide precursors. , 2000, Archives of insect biochemistry and physiology.

[45]  L. Schoofs,et al.  Peptidomics of the pars intercerebralis-corpus cardiacum complex of the migratory locust, Locusta migratoria. , 2001, European journal of biochemistry.

[46]  J. Huybrechts,et al.  New insights in Adipokinetic Hormone (AKH) precursor processing in Locusta migratoria obtained by capillary liquid chromatography-tandem mass spectrometry 1 1 Note: Web page available at http://www.kuleuven.ac.be/bio/fysio. , 2002, Peptides.

[47]  D. N. Perkins,et al.  Probability‐based protein identification by searching sequence databases using mass spectrometry data , 1999, Electrophoresis.

[48]  R. Kellner,et al.  Identification of two allatostatins from the cricket, Gryllus bimaculatus de Geer (Ensifera, Gryllidae): additional members of a family of neuropeptides inhibiting juvenile hormone biosynthesis , 1995, Regulatory Peptides.

[49]  L. Schoofs,et al.  Peptidomics of the Larval Drosophila melanogasterCentral Nervous System* , 2002, The Journal of Biological Chemistry.

[50]  J. Broeck.  Neuropeptides and their precursors in the fruitfly, Drosophila melanogaster☆ , 2001, Peptides.

[51]  M. Adams,et al.  Identification of Ecdysis-Triggering Hormone from an Epitracheal Endocrine System , 1996, Science.

[52]  J. Dow,et al.  Isolation and characterization of a leucokinin-like peptide of Drosophila melanogaster. , 1999, The Journal of experimental biology.

[53]  J. Broeck,et al.  Molecular cloning of the precursor cDNA for schistostatins, locust allatostatin-like peptides with myoinhibiting properties , 1996, Molecular and Cellular Endocrinology.

[54]  C. Slaughter,et al.  The fruitfly Drosophila melanogaster contains a novel charged adipokinetic-hormone-family peptide. , 1990, The Biochemical journal.

[55]  P. Verleyen,et al.  Comparison of the effects of pyrokinins and related peptides identified from arthropods on pupariation behaviour in flesh fly (Sarcophaga bullata) larvae (Diptera: Sarcophagidae). , 2004, Journal of insect physiology.

[56]  M. Moniatte,et al.  Differential display of peptides induced during the immune response of Drosophila: a matrix-assisted laser desorption ionization time-of-flight mass spectrometry study. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[57]  P. Taghert,et al.  Isolation and characterization of a Drosophila gene that encodes multiple neuropeptides related to Phe-Met-Arg-Phe-NH2 (FMRFamide). , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[58]  J. Rehfeld,et al.  The sulfakinins of the blowfly Calliphora vomitoria. Peptide isolation, gene cloning and expression studies. , 1995, European journal of biochemistry.

[59]  T. Mes,et al.  Adipokinetic peptide hormone content and biosynthesis during locust development , 1993, Peptides.

[60]  L. Schoofs,et al.  Expression of a novel neuropeptide, NVGTLARDFQLPIPNamide, in the larval and adult brain of Drosophila melanogaster , 2003, Journal of neurochemistry.

[61]  J. Yates,et al.  Large-scale analysis of the yeast proteome by multidimensional protein identification technology , 2001, Nature Biotechnology.

[62]  R. Feyereisen,et al.  Identification of an allatostatin from adult Diploptera punctata. , 1989, Biochemical and biophysical research communications.

[63]  J. Shabanowitz,et al.  Identity of a second type of allatostatin from cockroach brains: an octadecapeptide amide with a tyrosine-rich address sequence. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[64]  J. Veenstra.  Isolation and structure of corazonin, a cardioactive peptide from the American cockroach , 1989, FEBS letters.

[65]  D. Richter,et al.  Reverse physiology in Drosophila: identification of a novel allatostatin‐like neuropeptide and its cognate receptor structurally related to the mammalian somatostatin/galanin/opioid receptor family , 1999, The EMBO journal.

[66]  J. Truman,et al.  Isolation, characterization and expression of the eclosion hormone gene of Drosophila melanogaster. , 1993, European journal of biochemistry.