Fourier transform mass spectrometry: a powerful tool for toxin analysis.
The crude venom of Conus virgo was analyzed by Fourier transform mass spectrometry (FTMS) using both nano-electrospray ionization and MALDI. The analyses were performed directly on the crude venom, without chromatographic separation. The mass fingerprinting of the venom yielded 64 distinct molecular masses in the range 500-4500 Da with two major components at 1328.5142 and 1358.5592 Da. To facilitate the de novo sequencing of these compounds, the disulfide bonds of all components were reduced for the whole venom. The mass accuracy, resolution and sensitivity provided by FTMS were necessary to complete the sequencing of the two new peptides named ViVA and ViVB, that turned out to be conotoxins belonging to the T-superfamily, with the disulfide framework V. The peptides shared 80% similarity and as often observed for this class of compound, they were highly post-translationally modified: amidated C-terminus, pyroglutamic acid residue at the N-terminus and two disulfide bonds. Complementary online nano-LC-nano-ESI-FTMS experiments were undertaken. Among the 130 molecular masses found in the coupling experiments, only 45 were common with those obtained in the direct approach, which means that 21 compounds observed by nano-ESI-FTMS were not detected. This clearly shows that some discriminations against some classes of compounds occur when a chromatographic step is used before mass spectrometry.
Laboratoire des Mécanismes Réactionnels, UMR 7651 CNRS, Ecole Polytechnique, F-91128 Palaiseau, France.
Loïc
L
Quinton
Jean-Pierre
JP
Le Caër
Joëlle
J
Vinh
Nicolas
N
Gilles
Julia
J
Chamot-Rooke
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D020916:Conotoxins / Q000032:analysis*; D005583:Fourier Analysis; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010679:Philippines; D010720:Phosphines; D011499:Protein Processing, Post-Translational; D030162:Reptilian Proteins / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D020916:Conotoxins; D010720:Phosphines; D030162:Reptilian Proteins; C080938:tris(2-carboxyethyl)phosphine