title: Peptide fingerprinting of snake venoms by direct infusion nano-electrospray ionization mass spectrometry: potential use in venom identification and taxonomy.
abstract: Fingerprinting by mass spectrometry has been increasingly used to study venom variations and for taxonomic analyses based on venom components. Most of these studies have concentrated on components heavier than 3 kDa, but Bothrops snake venoms contain many biologically active peptides, principally C-type natriuretic peptides and bradykinin-potentiating peptides (BPPs). In this work, we have examined the peptide profile of Bothrops venoms (B. alternatus, B. erythromelas, B. insularis, B. jararaca, B. jararacussu, B. leucurus and B. moojeni) using direct infusion nano-electrospray ionization mass spectrometry (nano-ESI-MS) subjecting the data further to principal components analysis (PCA) to assess whether the peptide distributions are reliable in distinguishing the venoms. ESI-MS of a low molar mass fraction obtained by ultrafiltration of each venom (5 kDa nominal cutoff filters) revealed that the venoms have a variety of peptides in common but that each venom also contains taxonomic marker peptides not shared with other venoms. One BPP peptide, QGGWPRPGPEIPP, was found to be common to the seven Bothrops species examined. This peptide may represent a specific marker for this genus since it was not found in the venom of the South American rattlesnake, Crotalus durissus terrificus. PCA on the ESI-MS data reveals a close relationship between B. jararaca, B. jararacussu and B. moojeni venoms, with B. leucurus and B. erythromelas being more distant from these three; B. alternatus and B. insularis were also located distant from these five species, as was C. d. terrificus. These results agree partially with established phylogenetic relationships among these species and suggest that ESI-MS peptide fingerprinting of snake venoms coupled with PCA is a useful tool for identifying venoms and for taxonomic analyses.
authors.affiliation: Department of Pharmacology, Faculty of Medical Sciences, State University of Campinas (UNICAMP), Campinas, SP, Brazil. gustavo@brmass.com.br
authors.forename: Gustavo H M F
authors.initials: GH
authors.lastname: Souza
authors.forename: Rodrigo R
authors.initials: RR
authors.lastname: Catharino
authors.forename: Demian R
authors.initials: DR
authors.lastname: Ifa
authors.forename: Marcos N
authors.initials: MN
authors.lastname: Eberlin
authors.forename: Stephen
authors.initials: S
authors.lastname: Hyslop
mesh_terms: D005240:Feasibility Studies; D017022:Flow Injection Analysis / Q000379:methods*; D008832:Microchemistry / Q000379:methods*; D036103:Nanotechnology / Q000379:methods*; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000737:chemistry* / Q000145:classification; D012910:Snake Venoms / Q000737:chemistry* / Q000145:classification; D013048:Specimen Handling / Q000379:methods; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D010455:Peptides; D012910:Snake Venoms