Molecular and mass spectral identification of the broadly conserved decapod crustacean neuropeptide pQIRYHQCYFNPISCF: The first PISCF-allatostatin (Manduca sexta- or C-type allatostatin) from a non-insect

作者：Stemmler Elizabeth A^*; Bruns Emily A; Cashman Christopher R; Dickinson Patsy S; Christie Andrew E

来源：General and Comparative Endocrinology, 2010, 165(1): 1-10.

DOI：10.1016/j.ygcen.2009.05.010

• C-type allatostatin (C-AST)
• Manduca sexta-type allatostatin
• PISCF-allatostatin
• Expressed sequence tag (EST)
• Matrix-assisted laser desorption/ionization
• Fourier transform mass spectrometry (MALDI-FTMS) Neurchormone
• Neuropeptide
• Sustained off-resonance irradiation
• collision-induced dissociation (SORI-CID)

The PISCF-allatostatins (Manduca sexta- or C-type allatorstatins) are a family of pentadecapeptides characterized by a pyroglutamine blocked N-terminus, an unamidated-PISCF C-terminus, and a disulfide bridge between two internal Cys residues. Several isoforms of PISCF-AST are known, all from holometabolous insects. Using a combination of transcriptomics and mass spectrometry, we have identified the first PISCF-type peptides from a non-insect species. In silica analysis of crustacean ESTs identified several Litopenaeus vannamei (infraorder Penaeidea) transcripts encoding putative PISCF-AST precursors. Translation of these ESTs, with subsequent prediction of their putative post-translational processing, revealed the existence of as many as three PISCF-type peptides, including pQIRYHQCYFNPISCF (disulfide bridging between CyS7 and CyS14). Although none of the predicted isoforms was detected by mass spectrometry in L. vannamei, MALDI-FTMS mass profiling identified an m/z signal corresponding to pQIRYHQCYFNPISCF (disulfide bridge present) in neural tissue from 28 other decapods, which included members of six infra-orders (Stenopodidea, Astacidea, Thalassinidea, Achelata, Anomura and Brachyura). Further characterization of the peptide using SORI-CID and chemical derivatization/enzymatic digestion supported the theorized structure. In both the crab Cancer borealis and the lobster Homarus americanus, MALDI-based tissue surveys suggest that pQIRYHQCYFNPISCF is broadly distributed in the nervous system; it was also detected in the posterior midgut caecum. Collectively, our data show that members of the PISCF-AST family are not restricted to the holometabolous insects, but instead may be broadly conserved within the Pancrustacea. Moreover, our data suggest that one highly conserved PISCF-type peptide, pQIRYHQCYFN-PISCF, is present in decapod crustaceans, functioning as a brain-gut paracrine/hormone.

• 出版日期2010-1-1