The multi-component venom of the spider Cupiennius salei: A bioanalytical investigation applying different strategies

TRACHSEL, CHRISTIAN; SIEGEMUND, DOREENC; KÄMPFER, URS; KOPP, LUKAS; BÜHR, CLAUDIA; GROSSMANN, JONAS; LÜTHI, CHRISTOPH; MONICA CUNNINGHAM; NENTWIG, WOLFGANG; KUHN-NENTWIG, LUCIA; SCHÜRCH, STEFAN; SCHALLER, JOHANN

FEBS JOURNAL

WILEY-BLACKWELL PUBLISHING, INC

Lugar: Londres; Año: 2012 vol. 279 p. 2683 - 2694

1742-464X

The multicomponent venom of the spider Cupiennius salei was separatedby three different chromatographic strategies to facilitate subsequent analysisof peptidic venom components by tandem mass spectrometry (MALDITOF-MS and ESI-MS), Edman degradation and amino acid analysis: (a)desalting of the crude venom by RP-HPLC only, (b) chromatographic separationof the crude venom into 42 fractions by RP-HPLC, and (c) multidimensionalpurification of the crude venom by size exclusion and cationexchange chromatography and RP-HPLC. A total of 286 components wereidentified in the venom of C. salei by mass spectrometry and the sequenceof 49 new peptides was determined de novo by Edman degradation andtandem mass spectrometry; 30 were C-terminally amidated. The novel peptideswere assigned to two main groups: (a) short cationic peptides and (b)Cys-containing peptides with the inhibitor cystine knot motif. Bioinformaticsrevealed a limited number of substantial similarities, namely with thepeptides CpTx1 from the spider Cheiracantium punctorium and U3-ctenitoxin-Asp1a from the South American fishing spider (Ancylometes sp.) andwith sequences from a Lycosa singoriensis venom gland transcriptome analysis.The results clearly indicate that the quality of the data is stronglydependent on the chosen separation strategy. The combination of orthogonalanalytical methods efficiently excludes alkali ion and matrix adducts,provides indispensable information for an unambiguous identification ofisomasses, and results in the most comprehensive repertoire of peptidesidentified in the venom of C. salei so far.