Soluble expression, purification and functional identification of a disulfide-rich conotoxin derived from Conus litteratus.
Conotoxins are a diverse array of small peptides mostly with multiple disulfide bridges. These peptides become an increasing significant source of neuro-pharmacological probes and drugs as a result of the high selectivity for ion channels and receptors. Usually, the analogue of natural conotoxins is produced by means of chemical synthesis. Here, we present a simple and fast strategy of producing disulfide-rich conotoxins via recombinant expression. By fused with thioredoxin and His tag, a novel O-superfamily conotoxin lt7a was successfully expressed in Escherichia coli and purified, resulting in a high yield of recombinant lt7a about 6 mg/l. The purity of target protein is up to 95% as identified by HPLC results. Whole cell patch-clamp recording revealed that the new conotoxin blocked voltage-sensitive sodium channels in rat dorsal root ganglion neurons, indicating it might be a novel microO-conotoxin.
Pi
Canhui
C
State Key Laboratory of Biocontrol, Guangdong Province Key Laboratory of Therapeutic Functional Genes, Open Laboratory for Marine Functional Genomics, Department of Biochemistry, College of Life Sciences, Sun Yat-sen University, People's Republic of China.
Liu
Junliang
J
Wang
Lei
L
Jiang
Xiuhua
X
Liu
Yun
Y
Peng
Can
C
Chen
Shangwu
S
Xu
Anlong
A
D000818:Animals; D001483:Base Sequence; D020916:Conotoxins / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D052078:Conus Snail / Q000378:metabolism; D004220:Disulfides; D004594:Electrophysiology; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D005727:Ganglia, Spinal / Q000187:drug effects / Q000502:physiology*; D008297:Male; D008969:Molecular Sequence Data; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000494:pharmacology; D015222:Sodium Channels / Q000187:drug effects
D020916:Conotoxins; D004220:Disulfides; D011994:Recombinant Proteins; D015222:Sodium Channels