title: A novel conotoxin from Conus betulinus, kappa-BtX, unique in cysteine pattern and in function as a specific BK channel modulator.
abstract: A novel conotoxin, kappa-conotoxin (kappa-BtX), has been purified and characterized from the venom of a worm-hunting cone snail, Conus betulinus. The toxin, with four disulfide bonds, shares no sequence homology with any other conotoxins. Based on a partial amino acid sequence, its cDNA was cloned and sequenced. The deduced sequence consists of a 26-residue putative signal peptide, a 31-residue mature toxin, and a 13-residue extra peptide at the C terminus. The extra peptide is cleaved off by proteinase post-processing. All three Glu residues are gamma-carboxylated, one of the two Pro residues is hydroxylated at position 27, and its C-terminal residue is Pro-amidated. The monoisotopic mass of the toxin is 3569.0 Da. Electrophysiological experiments show that: 1) among voltage-gated channels, kappa-BtX is a specific modulator of K(+) channels; 2) among the K channels, kappa-BtX specifically up-modulates the Ca(2+)- and voltage-sensitive BK channels (252 +/- 47%); 3) its EC(50) is 0.7 nm with a single binding site (Hill = 0.88); 4) the time constant of wash-out is 8.3 s; and 5) kappa-BtX has no effect on single channel conductance, but increases the open probability of BK channels. It is concluded that kappa-BtX is a novel specific biotoxin against BK channels.
authors.affiliation: Research Institute of Pharmaceutical Chemistry, Beijing 102205, China.
authors.forename: Chong-Xu
authors.initials: CX
authors.lastname: Fan
authors.forename: Xiao-Ke
authors.initials: XK
authors.lastname: Chen
authors.forename: Chen
authors.initials: C
authors.lastname: Zhang
authors.forename: Li-Xiu
authors.initials: LX
authors.lastname: Wang
authors.forename: Kai-Lai
authors.initials: KL
authors.lastname: Duan
authors.forename: Lin-Lin
authors.initials: LL
authors.lastname: He
authors.forename: Ying
authors.initials: Y
authors.lastname: Cao
authors.forename: Shang-Yi
authors.initials: SY
authors.lastname: Liu
authors.forename: Ming-Nai
authors.initials: MN
authors.lastname: Zhong
authors.forename: Chris
authors.initials: C
authors.lastname: Ulens
authors.forename: Jan
authors.initials: J
authors.lastname: Tytgat
authors.forename: Ji-Sheng
authors.initials: JS
authors.lastname: Chen
authors.forename: Cheng-Wu
authors.initials: CW
authors.lastname: Chi
authors.forename: Zhuan
authors.initials: Z
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015220:Calcium Channels / Q000502:physiology; D002478:Cells, Cultured; D019439:Chromaffin Cells / Q000187:drug effects / Q000502:physiology; D002850:Chromatography, Gel; D020916:Conotoxins / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D051036:Large-Conductance Calcium-Activated Potassium Channels; D008969:Molecular Sequence Data; D008974:Mollusca; D015221:Potassium Channels / Q000502:physiology; D024681:Potassium Channels, Calcium-Activated / Q000037:antagonists & inhibitors*; D051381:Rats; D017208:Rats, Wistar; D015222:Sodium Channels / Q000502:physiology; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D015220:Calcium Channels; D020916:Conotoxins; D017931:DNA Primers; D018076:DNA, Complementary; D051036:Large-Conductance Calcium-Activated Potassium Channels; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D015222:Sodium Channels; C475056:kappa-conotoxin BtX