Nanospray analysis of the venom of the tarantula Theraphosa leblondi: a powerful method for direct venom mass fingerprinting and toxin sequencing |
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Authors: | Legros Christian Célérier Marie-Louise Henry Maud Guette Catherine |
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Affiliation: | UMR CNRS 8587, Laboratoire Analyse et Environnement, Institut des Sciences, Université d'Evry Val-d'Essonne, Bd. Fran?ois Mitterrand, 91024 Evry Cedex, France. |
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Abstract: | Mass spectrometric methods, including matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOFMS), on-line liquid chromatography/electrospray ionisation mass spectrometry (LC/ESI-MS), and nanospray ionisation/hybrid quadrupole time-of-flight mass spectrometry (nanoESI-QqTOFMS), were applied to characterize by mass fingerprinting the venom of the French Guyanese tarantula Theraphosa leblondi. Of these techniques direct nanoESI-QqTOFMS, which allowed the detection of 65 protonated molecules with high mass accuracy, appeared to give the best results. Three major peptides, TlTx1, TlTx2 and TlTx3, were sequenced using a combination of nanoESI-MS/MS and enzyme digestion/MS and MS/MS experiments. Each sequence was confirmed by automated Edman sequencing. In patch-clamp experiments these peptides were found to have a specific inhibitory effect on the voltage-dependent potassium channel, Kv4.2. |
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