Functional expression of α‐latrotoxin in baculovirus system

FEBS Letters

Volynski, Kirill E¹  Nosyreva, Elena D¹  Ushkaryov, Yuri A²  Grishin, Eugene V¹ 

[1] Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow 117871, Russia;Department of Biochemistry, Imperial College, Exhibition Road, London SW7 2AY, UK

关键词: α-Latrotoxin;    Baculovirus;    Expression;    Latrophilin;    Neurexin;   

DOI  :  10.1016/S0014-5793(98)01624-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown