Extraction and partial characterization of venom from the Colombian spider Pamphobeteus aff. nigricolor (Aranae:Theraphosidae)

• Published in: Toxicon (Oxford) 2013-12, Vol.76, p.301-309
• Main Authors: Estrada-Gomez, Sebastian, Vargas Muñoz, Leidy Johana, Quintana Castillo, Juan C.
• Format: Article
• Language: English
• Subjects: anesthesia; Animals; Araneae; calcium channels; Channels; Chemical Fractionation; Chromatography, High Pressure Liquid; Chromatography, Reverse-Phase; Colombia; Density; Drying; electrophoresis; Electrophoresis, Polyacrylamide Gel; Extraction; Female; Females; hemolysis; humidity; Male; Males; Media; molecular weight; Pamphobeteus nigricolor; Peptides; protein content; proteolysis; reversed-phase high performance liquid chromatography; RP-HPLC; Sex Factors; Spider Venoms - chemistry; Spider Venoms - isolation & purification; Spiders - chemistry; Theraphosidae; Venom extraction; venoms; viscosity
• ISSN: 0041-0101; 1879-3150
• DOI: 10.1016/j.toxicon.2013.10.014

We report the first studies of characterization and extraction of the Pamphobeteus aff. nigricolor (Pocock, 1901) (Aranae:Theraphosidae) venom done in Colombia using the electro-stimulation technique previous anesthesia with isofluorane. After each extraction process, a low viscosity, colorless venom was obtained. This venom showed a 1.01 mg/μl density and a pH of 5. The humidity percentage did not show a significance difference between males and females (P > 0.05) with a general media of 77.49 ± 1.74%. In all cases the venom yielded was variable between males and females, with a media of 22.45 ± 5.17 mg (wet weight) and 4.58 ± 0.94 mg (dry weigh), obtaining larger amounts in females, 28.34 ± 7.49 mg and 5.69 ± 1.36 (wet and dry weight respectively). Venom showed a hemolytic activity dependent of enzymatic active phospholipase and neither coagulant nor proteolytic activities were observed. Electrophoretic profile showed a main protein content with a molecular mass below 14 kDa. RP-HPLC venom profile revealed a difference among male and female venom's content where 17 and 21 main fractions were obtained respectively. Three peptides, Theraphotoxin-Pn1a, Theraphotoxin-Pn1b and Theraphotoxin-Pn2a, were identified using HPLC-nESI-MS/MS. These peptides showed a high identity with other peptides found on Theraphosides which are proved to affect voltage-gated calcium channels. •Venom characteristics from the Colombian Theraphosidae Pamphobeteus nigricolor are reported for first time.•Venoms properties agree with other Theraphosidae genus venoms.•Biochemical profile may indicate the presence of cytolytic peptides.•Electrophoretic and chromatographic profile indicates presence of low molecular mass components.