Cloning of neuropeptide-like precursor 1 in the gray flesh fly and peptide identification and expression.
The neuropeptide-like precursor 1 (NPLP1) was first identified in a peptidomics experiment on Drosophila melanogaster. Limited data on this novel neuropeptide precursor suggest a role in the regulation of ecdysis in holometabolous larvae. In this study, we characterized the NPLP1 precursor in the gray flesh fly, Neobellieria bullata, which is an excellent model for physiological assays and hence to discover the role of the NPLP1 peptides. Antisera against three of the D. melanogaster NPLP1 neuropeptides stained an identical set of neurons in the central nervous system of N. bullata compared to D. melanogaster. A novel approach was applied to identify the N. bullata NPLP1 orthologs. Using a combination of affinity chromatography, mass spectrometry, cDNA cloning and RACE experiments, we obtained almost the complete coding sequence of the NPLP1 mRNA. Three encoded NPLP1 peptides were identified in central nervous system extracts by mass spectrometry. Neither doses of 25-250pmol of synthetic Neb-MGYamide and Neb-PQNamide peptides, nor the NPLP1 antisera did affect the speed of retraction, contraction and tanning in the pupariation bioassay.
Verleyen
Peter
P
Research Group Functional Genomics and Proteomics, K.U. Leuven, Naamsestraat 59, B-3000 Leuven, Belgium. peter.verleyen@bio.kuleuven.be
Chen
Xi
X
Baron
Szilvia
S
Preumont
Alice
A
Hua
Yue Jin
YJ
Schoofs
Liliane
L
Clynen
Elke
E
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003001:Cloning, Molecular; D004175:Diptera / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009479:Neuropeptides / Q000235:genetics* / Q000302:isolation & purification; D011679:Pupa / Q000502:physiology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D029721:Drosophila Proteins; D009479:Neuropeptides; C518480:Nplp1 protein, Drosophila