title: High throughput screening of bradykinin-potentiating peptides in Bothrops moojeni snake venom using precursor ion mass spectrometry.
abstract: Snake venoms are known to be an extensive source of bioactive peptides. Bradykinin-potentiating peptides (BPPs) are inhibitors of the angiotensin-converting enzyme that have already been identified in the venom of many snake, scorpion, spider and batrachian species. Their most characteristic structural features are an invariable N-terminal pyroglutamate residue (pGlu or Z) and two consecutive proline residues at the C-terminus. Fragmentation of BPPs by collision-induced dissociation during electrospray tandem mass spectrometry analysis (ESI-MS/MS) generates a predominant signal at m/z 213.1 corresponding to the y-ion of the terminal Pro-Pro fragment. In addition, signals at m/z 226.1 and 240.1 that correspond to the b ions of the N-terminus pGlu-Asn and pGlu-Lys, respectively, can often be observed. Based on these structural determinants, the present work describes an original methodology for the discovery of BPPs in natural extracts using liquid chromatography coupled to ESI-MS/MS operated in precursor ion-scan mode. The venom of the Bothrops moojeni snake was used as a model and the methodology was applied for subsequent structural analysis of the identified precursors by tandem mass spectrometry on quadrupole-time-of-flight (Q-TOF) and matrix-assisted laser-desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF-MS/MS) instruments. More than 40 peptides below 2500 Da could be detected, among them 20 were shown to belong to the BPP-like family including the related tripeptides pGlu-Lys-Trp and pGlu-Asn-Trp. A total of 15 new sequences have been identified using this approach.
authors.affiliation: Atheris Laboratories, Case Postale 314, CH-1233 Bernex, Geneva, Switzerland.
authors.forename: Laure
authors.initials: L
authors.lastname: Menin
authors.forename: Anna
authors.initials: A
authors.lastname: Perchuć
authors.forename: Philippe
authors.initials: P
authors.lastname: Favreau
authors.forename: Frédéric
authors.initials: F
authors.lastname: Perret
authors.forename: Sophie
authors.initials: S
authors.lastname: Michalet
authors.forename: Reto
authors.initials: R
authors.lastname: Schöni
authors.forename: Marianne
authors.initials: M
authors.lastname: Wilmer
authors.forename: Reto
authors.initials: R
authors.lastname: Stöcklin
mesh_terms: D000818:Animals; D017837:Bothrops / Q000378:metabolism*; D001920:Bradykinin / Q000737:chemistry*; D003435:Crotalid Venoms / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods*; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping; D010455:Peptides / Q000737:chemistry*; D011761:Pyrrolidonecarboxylic Acid / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry
chemical_list: D003435:Crotalid Venoms; D010446:Peptide Fragments; D010455:Peptides; D001920:Bradykinin; D011761:Pyrrolidonecarboxylic Acid