Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry and high‐performance liquid chromatography study of quantitative and qualitative variation in tarantula spider venoms

Abstract

Animal venoms are important sources of novel pharmacological tools, useful in biochemical characterization of their receptors. Venom quality control, batch-to-batch homogeneity and high reproducibility of venom fractionation and toxin purification are crucial issues for biochemical and pharmacological studies. To address these issues, a study of the variability of tarantula spider venom samples was undertaken. Venom profiles of samples collected from individuals of different age and sex, and from sibling spiders of the same species, were generated by high-performance liquid chromatography (HPLC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and analyzed to assess venom variability and method accuracy. Sex-linked venom variation was studied on eight species. Clear qualitative differences were observed for six out of eight species, as well as quantitative differences. Age-related variation studied in Poecilotheria rufilata showed essentially age-related quantitative differences between adults of both sexes and immature juveniles. The venoms of nine siblings and three wild-collected Pterinochilus murinus were studied for individual variation, showing only very minor quantitative differences. On the same samples, the quality of MALDI-TOFMS venom fingerprinting was demonstrated to be highly reproducible. Our results show that tarantula venom peptide fingerprinting is a highly reliable identification method, that pooled batches of venom from several animals can be used for venom purification, that venom composition does not appear to be qualitatively related to ontogenesis in the spiders studied, and that qualitative sex-linked variation occurs across most species and may be important in activity studies.

Intraspecific variations in the venom peptidome of the ant Odontomachus haematodus (Formicidae: Ponerinae) from French Guiana

“…However, intraspecific variations can also occur as shown recently in Dinoponera quadriceps for which only 48 peptides were shared between colonies out of the more than 300 peptides found in total (Cologna et al 2013). Also, intraspecific variations in venom composition have been observed in snakes, scorpions, tarantulas and cone snails, this variation being linked to geographical distribution (Núñez et al 2009;Shashidharamurthy et al 2002), age (Escoubas et al 2002) or sex (Escoubas et al 1997;Herzig and Hodgson 2009;Herzig et al 2002;Herzig et al 2008). However, it remains unclear whether such variation is a common denominator for all venomous animals or is restricted to some taxa or species.…”

Intraspecific variation of venom injected by fish-hunting Conussnails

“…Significantly less dramatic variations in the peptide complement of injected venoms have been reported for several other organisms, including snakes (Chippaux et al, 1991;Creer et al, 2003;Daltry et al, 1996a,b;Francischetti et al, 2000;MacKessy et al, 2003;Monteiro et al, 1998a,b), spiders (Binford, 2001;Cristina de Oliveira et al, 1999;Escoubas et al, 2002) The α-, ω-conotoxins and s4a have all been previously isolated from the DV and characterized. Underlined peptides are those that are putatively identified in this work by mass matching to published cDNA sequences with predicted modifications.…”

“…Sequence s4b is not included in this and Her, 2000). Variation of the peptide profiles of these other venomous animals has been associated with sex (Binford, 2001;Cristina de Oliveira et al, 1999;Escoubas et al, 2002), diet (Daltry et al, 1996b), age (Escoubas et al, 2002), geography (Binford, 2001;Creer et al, 2003), season (Monteiro et al, 1998b) and venom regeneration time (Pimenta et al, 2003). Studies controlling many of these factors have still observed venom variation, implying that intraspecific differences can be a result of genetic as well as environmental factors (Daltry et al, 1996a;Francischetti et al, 2000;Kalapothakis and Chavez-Olortegui, 1997;Monteiro et al, 1998a,b).…”

Isolation And Some Characterizations Of A Glycosylated Fibrinolytic Enzyme Of Earthworm, Eisenia Fetida

“…according to Escoubas et al [15]. All experiments were performed using the saturated α-cyano-4-hydroxycinnamic acid (CCA), dissolved in 50% (v/v) solution of acetonitrile and aqueous 0.2% TFA, as the matrix.…”