title: Recombinant Expression, Functional Characterization of Two Scorpion Venom Toxins with Three Disulfide Bridges from the Chinese Scorpion Buthus martensii Karsch.
abstract: Scorpion venom contains a large variety of biologically active peptides. However, most of these peptides have not been identified and characterized. Peptides with three disulfide bridges, existing in the scorpion venom, have not been studied in detail and have been poorly characterized until now. Here, we report the recombinant expression and functional characterization of two kinds of venom peptides (BmKBTx and BmNaL-3SS2) with three disulfide bridges. This study adopted an effective Escherichia coli system. The genes for BmKBTx and BmNaL-3SS2 were obtained by polymerase chain reaction and cloned to the pSYPU-1b vector. After expression and purification, the two recombinant proteins were subjected to an analgesic activity assay in mice and whole-cell patchclamp recording of hNav1.7-CHO cell lines. Functional tests showed that BmKBTx and BmNaL- 3SS2 have analgesic activity in mice and can interact with the hNav1.7 subtype of the voltage-gated sodium channel (VGSC). Scorpion venom is rich in bioactive proteins, but most of their functions are unknown to us. This study has increased our knowledge of these novel disulfide-bridged peptides (DBPs) and their biological activities.
authors.forename: Shengguo
authors.initials: S
authors.lastname: Lin
authors.forename: Xuelin
authors.initials: X
authors.lastname: Wang
authors.forename: Xueyao
authors.initials: X
authors.lastname: Hu
authors.forename: Yongshan
authors.initials: Y
authors.lastname: Zhao
authors.forename: Mingyi
authors.initials: M
authors.lastname: Zhao
authors.affiliation: School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University, Shenyang, Liaoning Province 110016, China.
authors.forename: Jinghai
authors.initials: J
authors.lastname: Zhang
authors.affiliation: School of Medical Devices, Shenyang Pharmaceutical University, Benxi, Liaoning Province 117004, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Cui
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000595:Amino Acid Sequence; D000700:Analgesics / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D002681:China; D003001:Cloning, Molecular; D003412:Cricetulus; D004220:Disulfides / Q000737:chemistry; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D008297:Male; D051379:Mice; D062556:NAV1.7 Voltage-Gated Sodium Channel / Q000235:genetics* / Q000378:metabolism; D018408:Patch-Clamp Techniques; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D012604:Scorpion Venoms / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D012605:Scorpions / Q000737:chemistry / Q000502:physiology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D000700:Analgesics; D004220:Disulfides; C418407:KBT toxin, Buthus martensii; D062556:NAV1.7 Voltage-Gated Sodium Channel; D011994:Recombinant Proteins; C486866:SCN9A protein, human; D012604:Scorpion Venoms
keywords: BmKBTx; BmNaL-3SS2; analgesic activity; disulfide bridge; electrophysiology; hNav1.7; recombinant expression