title: BmP09, a "long chain" scorpion peptide blocker of BK channels.
abstract: A novel "long chain" toxin BmP09 has been purified and characterized from the venom of the Chinese scorpion Buthus martensi Karsch. The toxin BmP09 is composed of 66 amino acid residues, including eight cysteines, with a mass of 7721.0 Da. Compared with the B. martensi Karsch AS-1 as a Na(+) channel blocker (7704.8 Da), the BmP09 has an exclusive difference in sequence by an oxidative modification at the C terminus. The sulfoxide Met-66 at the C terminus brought the peptide a dramatic switch from a Na(+) channel blocker toaK(+) channel blocker. Upon probing the targets of the toxin BmP09 on the isolated mouse adrenal medulla chromaffin cells, where a variety of ion channels coexists, we found that the toxin BmP09 specifically blocked large conductance Ca(2+)- and voltage-dependent K(+) channels (BK) but not Na(+) channels at a range of 100 nm concentration. This was further confirmed by blocking directly the BK channels encoded with mSlo1 alpha-subunits in Xenopus oocytes. The half-maximum concentration EC(50) of BmP09 was 27 nm, and the Hill coefficient was 1.8. In outside-out patches, the 100 nm BmP09 reduced approximately 70% currents of BK channels without affecting the single-channel conductance. In comparison with the "short chain" scorpion peptide toxins such as Charybdotoxin, the toxin BmP09 behaves much better in specificity and reversibility, and thus it will be a more efficient tool for studying BK channels. A three-dimensional simulation between a BmP09 toxin and an mSlo channel shows that the Lys-41 in BmP09 lies at the center of the interface and plugs into the entrance of the channel pore. The stable binding between the toxin BmP09 and the BK channel is favored by aromatic pi -pi interactions around the center.
authors.affiliation: Institute of Biochemistry and Biophysics, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei 430074, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Yao
authors.forename: Xiang
authors.initials: X
authors.lastname: Chen
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhou
authors.forename: Lijun
authors.initials: L
authors.lastname: Yao
authors.forename: Gong
authors.initials: G
authors.lastname: Wu
authors.forename: Xiaoke
authors.initials: X
authors.lastname: Chen
authors.forename: Naixia
authors.initials: N
authors.lastname: Zhang
authors.forename: Zhuan
authors.initials: Z
authors.lastname: Zhou
authors.forename: Tao
authors.initials: T
authors.lastname: Xu
authors.forename: Houming
authors.initials: H
authors.lastname: Wu
authors.forename: Jiuping
authors.initials: J
authors.lastname: Ding
mesh_terms: D000313:Adrenal Medulla / Q000166:cytology; D000595:Amino Acid Sequence; D000818:Animals; D018999:Charybdotoxin / Q000737:chemistry; D002845:Chromatography; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D006860:Hydrogen Bonding; D007700:Kinetics; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008239:Lysine / Q000737:chemistry; D008715:Methionine / Q000737:chemistry; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009865:Oocytes / Q000378:metabolism; D018408:Patch-Clamp Techniques; D010455:Peptides / Q000737:chemistry*; D024681:Potassium Channels, Calcium-Activated / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D012604:Scorpion Venoms / Q000737:chemistry* / Q000494:pharmacology*; D012605:Scorpions; D017386:Sequence Homology, Amino Acid; D012964:Sodium / Q000737:chemistry; D013454:Sulfoxides / Q000737:chemistry; D013997:Time Factors; D014981:Xenopus
chemical_list: D004220:Disulfides; D051036:Large-Conductance Calcium-Activated Potassium Channels; C500799:P09 protein, Buthus martensi; D010455:Peptides; D024681:Potassium Channels, Calcium-Activated; D012604:Scorpion Venoms; D013454:Sulfoxides; D018999:Charybdotoxin; D012964:Sodium; D008715:Methionine; D008239:Lysine; D003545:Cysteine