title: Characterization of endoproteolytic processing of dynorphins by proprotein convertases using mouse spinal cord S9 fractions and mass spectrometry.
abstract: Dynorphins are important neuropeptides with a central role in nociception and pain alleviation. Many mechanisms regulate endogenous dynorphin concentrations, including proteolysis. Proprotein convertases (PCs) are widely expressed in the central nervous system and specifically cleave at C-terminal of either a pair of basic amino acids, or a single basic residue. The proteolysis control of endogenous big dynorphin (BDyn) and dynorphin A (Dyn A) levels has a profound impact on pain perception and the role of PCs remain unclear. The objective of this study was to decipher the role of PC1 and PC2 in the proteolysis control of BDyn and Dyn A levels using cellular fractions of spinal cords from wild-type (WT), PC1(-/+) and PC2(-/+) animals and mass spectrometry. Our results clearly demonstrate that both PC1 and PC2 are involved in the proteolysis regulation of BDyn and Dyn A with a more important role for PC1. C-terminal processing of BDyn generates specific peptide fragments dynorphin 1-19, dynorphin 1-13, dynorphin 1-11 and dynorphin 1-7, and C-terminal processing of Dyn A generates dynorphin 1-13, dynorphin 1-11 and dynorphin 1-7, all these peptide fragments are associated with PC1 or PC2 processing. Moreover, the proteolysis of BDyn leads to the formation of Dyn A and Leu-Enk, two important opioid peptides. The rate of formation of both is significantly reduced in cellular fractions of spinal cord mutant mice. As a consequence, even the partial inhibition of PC1 or PC2 may impair the endogenous opioid system.
authors.affiliation: Groupe de Recherche en Pharmacologie Animal du Québec (GREPAQ), Faculté de Médecine Vétérinaire, Département de Biomédecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada.
authors.forename: Alberto Ruiz
authors.initials: AR
authors.lastname: Orduna
authors.affiliation: Groupe de Recherche en Pharmacologie Animal du Québec (GREPAQ), Faculté de Médecine Vétérinaire, Département de Biomédecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada. Electronic address: francis.beaudry@umontreal.ca.
authors.forename: Francis
authors.initials: F
authors.lastname: Beaudry
mesh_terms: D000818:Animals; D004399:Dynorphins / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D013058:Mass Spectrometry; D055442:Metabolome; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010447:Peptide Hydrolases; D045664:Proprotein Convertase 1 / Q000235:genetics / Q000378:metabolism*; D045706:Proprotein Convertase 2 / Q000235:genetics / Q000378:metabolism*; D013116:Spinal Cord / Q000201:enzymology*
chemical_list: D004399:Dynorphins; D010447:Peptide Hydrolases; D045664:Proprotein Convertase 1; D045706:Proprotein Convertase 2
keywords: Dynorphin A; Dynorphins; Mass spectrometry; Opioid peptides; Proprotein convertases; Proteolysis; Spinal cords