Expression, renaturation and functional analysis of an excitatory insect-specific toxin from scorpion Buthus martensii Karsch.
The cDNA of BmK IT-AP, an excitatory insect toxin from the scorpion Buthus martensi Karsch that has an analgesic effect on mammalian cells, was expressed in E. coli in the form of an inclusion body. Following denaturation and reduction, the recombinant protein was renatured and purified by liquid chromatography. The authenticity of the recombinant product was confirmed by bioassay and its electrophysiological effect on insect sodium channel.
Li
Chong
C
Center for Structural and Molecular Biology, Institute of Biophysics, Chinese Academy of Science, Beijing 100101, People's Republic of China.
Liu
Wei
W
Bossmans
Frank
F
Zhu
Rong-Huan
RH
Tytgat
Jan
J
Wang
Da-Cheng
DC
D000818:Animals; D004594:Electrophysiology; D015870:Gene Expression; D007814:Larva / Q000187:drug effects; D009036:Moths / Q000187:drug effects; D009865:Oocytes; D018408:Patch-Clamp Techniques; D020673:Protein Renaturation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000633:toxicity; D012604:Scorpion Venoms / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000633:toxicity*; D012605:Scorpions / Q000737:chemistry* / Q000235:genetics; D015222:Sodium Channels / Q000235:genetics / Q000378:metabolism; D014982:Xenopus laevis
D011994:Recombinant Proteins; D012604:Scorpion Venoms; D015222:Sodium Channels