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Nanospray analysis of the venom of the tarantula Theraphosa leblondi : a powerful method for direct venom mass fingerprinting and toxin sequencing
Author(s) -
Legros Christian,
Célérier MarieLouise,
Henry Maud,
Guette Catherine
Publication year - 2004
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.1442
Subject(s) - chemistry , chromatography , mass spectrometry , venom , edman degradation , time of flight mass spectrometry , electrospray ionization , electrospray , ionization , peptide sequence , biochemistry , ion , organic chemistry , gene
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. Copyright © 2004 John Wiley & Sons, Ltd.