Abstract
To facilitate the study of the mechanism of α-latrotoxin action, it is necessary to create a biologically active recombinant toxin. Mature α-latrotoxin is naturally produced by post-translational cleavage, probably at two furin sites located at the N- and C-termini of the precursor. A recombinant baculovirus has now been constructed, which encodes the melittin signal peptide fused to the 130-kDa mature toxin between the furin sites. Insect cells, infected with this baculovirus, secreted recombinant α-latrotoxin. This was partially purified and proved indistinguishable from the natural toxin with respect to its molecular mass, immunostaining, toxicity to mice, binding to α-latrotoxin receptors (latrophilin or neurexin Iα) and electrophysiological recording in the mouse diaphragm. The successful expression of recombinant α-latrotoxin permits mutational analysis of the toxin. Copyright (C) 1999 Federation of European Biochemical Societies.
Original language | English (US) |
---|---|
Pages (from-to) | 25-28 |
Number of pages | 4 |
Journal | FEBS Letters |
Volume | 442 |
Issue number | 1 |
DOIs | |
State | Published - Jan 8 1999 |
Keywords
- Baculovirus
- Expression
- Latrophilin
- Neurexin
- α-Latrotoxin
ASJC Scopus subject areas
- Biophysics
- Structural Biology
- Biochemistry
- Molecular Biology
- Genetics
- Cell Biology