title: Structure and orientation of a voltage-sensor toxin in lipid membranes.
abstract: Amphipathic protein toxins from tarantula venom inhibit voltage-activated potassium (Kv) channels by binding to a critical helix-turn-helix motif termed the voltage sensor paddle. Although these toxins partition into membranes to bind the paddle motif, their structure and orientation within the membrane are unknown. We investigated the interaction of a tarantula toxin named SGTx with membranes using both fluorescence and NMR spectroscopy. Depth-dependent fluorescence-quenching experiments with brominated lipids suggest that Trp30 in SGTx is positioned approximately 9 A from the center of the bilayer. NMR spectra reveal that the inhibitor cystine knot structure of the toxin does not radically change upon membrane partitioning. Transferred cross-saturation NMR experiments indicate that the toxin's hydrophobic protrusion contacts the hydrophobic core of the membrane, whereas most surrounding polar residues remain at interfacial regions of the bilayer. The inferred orientation of the toxin reveals a twofold symmetry in the arrangement of basic and hydrophobic residues, a feature that is conserved among tarantula toxins. These results have important implications for regions of the toxin involved in recognizing membranes and voltage-sensor paddles, and for the mechanisms by which tarantula toxins alter the activity of different types of ion channels.
authors.affiliation: Department of Life Science, Gwangju Institute of Science and Technology, Gwangju, Korea.
authors.forename: Hyun Ho
authors.initials: HH
authors.lastname: Jung
authors.forename: Hoi Jong
authors.initials: HJ
authors.lastname: Jung
authors.forename: Mirela
authors.initials: M
authors.lastname: Milescu
authors.forename: Chul Won
authors.initials: CW
authors.lastname: Lee
authors.forename: Seungkyu
authors.initials: S
authors.lastname: Lee
authors.forename: Ju Yeon
authors.initials: JY
authors.lastname: Lee
authors.forename: Young-Jae
authors.initials: YJ
authors.lastname: Eu
authors.forename: Ha Hyung
authors.initials: HH
authors.lastname: Kim
authors.forename: Kenton J
authors.initials: KJ
authors.lastname: Swartz
authors.forename: Jae Il
authors.initials: JI
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004252:DNA Mutational Analysis; D005453:Fluorescence; D015640:Ion Channel Gating; D008563:Membrane Lipids / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010713:Phosphatidylcholines / Q000737:chemistry; D011485:Protein Binding; D017433:Protein Structure, Secondary; D013057:Spectrum Analysis; D013111:Spider Venoms / Q000737:chemistry*; D014364:Tryptophan / Q000378:metabolism
chemical_list: D008563:Membrane Lipids; D010713:Phosphatidylcholines; D013111:Spider Venoms; D014364:Tryptophan