title: Mass spectrometry-assisted confirmation of the inability of dipeptidyl peptidase-4 to cleave goldfish peptide YY(1-36) and the lack of anorexigenic effects of peptide YY(3-36) in goldfish (Carassius auratus).
abstract: Dipeptidyl peptidase-4 (DPP4) is a serine protease of great interest because it has been shown to modulate the activity of several peptidergic factors including peptide YY (PYY) and glucagon-like peptide-1/2. While PYY(1-36) is orexigenic in mammals, PYY(3-36) recently garnered interest as a potent anorexigen. In silico phylogenetic analysis found that the DPP4 cleavage sites are absent in fish PYY sequences. However, no studies were conducted to show that indeed PYY(3-36) is not produced by DPP4 in fish. If DPP4 does not cleave PYY(1-36), is PYY(3-36) an anorexigen in fish? The objectives of this research were to (1) test whether DPP4 cleaves goldfish PYY(1-36) and (2) determine whether PYY(3-36) is an anorexigen in goldfish. First, we identified the highly conserved catalytic region of DPP4 in goldfish. Abundant expression of DPP4 mRNA was found within the gastrointestinal tract. We also report the first MALDI-MS cleavage analysis of DPP4 effects on PYY(1-36) in a non-mammalian vertebrate. Our novel results indicate that DPP4 is unable to cleave goldfish PYY(1-36) to PYY(3-36) in vitro. It also confirms a previously held hypothesis that DPP4 is unable to cleave fish PYY(1-36) that contains N-terminal proline-proline residues. PYY(3-36) had no effects on food intake of goldfish. The appetite inhibitory effects of intraperitoneal and intracerebroventricular injections of 10 ng/g body weight gfPYY(1-36) were abolished by coinjections of BIBP3226, a Y1 receptor antagonist. These results are significant because it shows the lack of generation of endogenous PYY(3-36) and its anorectic effects in goldfish.
authors.affiliation: Department of Biology, York University, Toronto, ON, Canada.
authors.forename: R
authors.initials: R
authors.lastname: Gonzalez
authors.affiliation: Laboratory of Integrative Neuroendocrinology, Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK, S7N 5B4, Canada. suraj.unniappan@usask.ca.
authors.forename: S
authors.initials: S
authors.lastname: Unniappan
mesh_terms: D000818:Animals; D018819:Dipeptidyl Peptidase 4 / Q000235:genetics / Q000378:metabolism*; D004435:Eating / Q000187:drug effects; D005260:Female; D029941:Fish Proteins / Q000378:metabolism* / Q000494:pharmacology; D041981:Gastrointestinal Tract / Q000378:metabolism; D006054:Goldfish; D008297:Male; D013058:Mass Spectrometry; D010446:Peptide Fragments / Q000378:metabolism* / Q000494:pharmacology; D019894:Peptide YY / Q000378:metabolism* / Q000494:pharmacology; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism
chemical_list: D029941:Fish Proteins; D010446:Peptide Fragments; D012333:RNA, Messenger; D019894:Peptide YY; C061785:peptide YY (3-36); D018819:Dipeptidyl Peptidase 4; C514549:peptide YY (1-36)
keywords: Brain; Dipeptidyl peptidase-4; Fish; Food intake; Mass spectrometry; Peptide YY