title: Extraction and partial characterization of venom from the Colombian spider Pamphobeteus aff. nigricolor (Aranae:Theraphosidae).
abstract: 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.
authors.affiliation: Facultad de Química Farmacéutica, Universidad de Antioquia UdeA, Medellín, Colombia; Programa de Ofidismo/Escorpionismo, Facultad de Química Farmacéutica, Universidad de Antioquia UdeA, Medellín, Colombia. Electronic address: sebastian.estrada@siu.udea.edu.co.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Estrada-Gomez
authors.forename: Leidy Johana
authors.initials: LJ
authors.lastname: Vargas Muñoz
authors.forename: Juan C
authors.initials: JC
authors.lastname: Quintana Castillo
mesh_terms: D000818:Animals; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid; D056148:Chromatography, Reverse-Phase; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D008297:Male; D012737:Sex Factors; D013111:Spider Venoms / Q000737:chemistry* / Q000302:isolation & purification; D013112:Spiders / Q000737:chemistry*
chemical_list: D013111:Spider Venoms
keywords: Pamphobeteus nigricolor; RP-HPLC; Theraphosidae; Venom extraction