A SPIDER TOXIN BINDING PROTEIN FROM BOVINE BRAIN : ITS PURIFICATION AND N-TERMINAL AMINO ACID SEQUENCE DETERMINATION

Anwar HOSSAIN; Ken'ichi HAGIWARA; Nobufumi KAWAI; Terumi NAKAJIMA

doi:10.1248/cpb.39.3079

Anwar HOSSAIN, Ken'ichi HAGIWARA, Nobufumi KAWAI, Terumi NAKAJIMA

Author information

Keywords: spider toxin binding protein, glutamate receptor, 1-naphthylacetyl spermine affinity chromatography, N-terminal amino acid analysis, JSTX-3, excitatory postsynaptic potential

JOURNAL FREE ACCESS

1991 Volume 39 Issue 11 Pages 3079-3081

DOI https://doi.org/10.1248/cpb.39.3079

Details

Abstract

A 60kDa spider toxin binding protein from bovine brain was solubilized with digitonin and purified up to 5800-folds over starting crude homogenate. The purification procedure entailed DEAE-cellulose, concanavalin-A affinity, 1-naphthylacetyl spermine affinity and high performance liquid chromatography. The purified protein owned a very high affinity for ligand ¹²⁵I-JSTX-3 binding Kd 15.6nM and Bmax 6.5nM. The amino acid composition of the protein was determined. The N-terminal amino acid sequence analysis yielded a unique sequence : NH₂-X-Pro-X-Val-Tyr-Phe-Lys-Glu-Gln-Phe-Leu-Asp-Gly-Asp-X-

Corresponding author

Register with J-STAGE for free!