title: A single-step purification of bothropstoxin-1.
abstract: Bothrops venoms are complex mixtures of components with a wide range of biological activities. Among these substances, myotoxins have been investigated by several groups. Bothropstoxin-1 (Bthtx-1) is a phospholipase A2-like basic myotoxin from Bothrops jararacussu. The purification of this component involves two chromatographic steps. Although providing a pure material, the association of these two steps is time consuming and a single-step method using high performance chromatography media would be useful. In the present study, we describe a single-step purification method for Bthtx-1. Bothrops jararacussu venom was dissolved in 1 ml buffer. After centrifugation, the supernatant was injected into a Resource-S cation exchange column connected to an FPLC system and eluted with a linear salt gradient. The complete procedure took 20 min, representing a considerable time gain when compared to a previously described method (Homsi-Brandenburgo MI et al. (1988) Toxicon, 26: 615-627). Bthtx-1 purity and identity, assessed by SDS-PAGE and N-terminal sequencing, resulted in a single band with a molecular mass of about 14 kDa and the expected sequence of the first 5 residues, S-L-F-E-L. Although the amount of protein purified after each run is lower than in the previously described method, we believe that this method may be useful for small-scale purifications.
authors.affiliation: IPEN/CNEN SP, São Paulo, Brasil. pspencer@net.ipen.br
authors.forename: P J
authors.initials: PJ
authors.lastname: Spencer
authors.forename: S D
authors.initials: SD
authors.lastname: Aird
authors.forename: M
authors.initials: M
authors.lastname: Boni-Mitake
authors.forename: N
authors.initials: N
authors.lastname: Nascimento
authors.forename: J R
authors.initials: JR
authors.lastname: Rogero
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003435:Crotalid Venoms / Q000302:isolation & purification*
chemical_list: D003435:Crotalid Venoms; C078097:bothropstoxin