Solution structure of the potassium channel inhibitor agitoxin 2: Caliper for probing channel geometry
Open Access
- 1 August 1995
- journal article
- research article
- Published by Wiley in Protein Science
- Vol. 4 (8), 1478-1489
- https://doi.org/10.1002/pro.5560040805
Abstract
The structure of the potassium channel blocker agitoxin 2 was solved by solution NMR methods. The structure consists of a triple‐stranded antiparallel β‐sheet and a single helix covering one face of the β‐sheet. The cysteine side chains connecting the β‐sheet and the helix form the core of the molecule. One edge of the β‐sheet and the adjacent face of the helix form the interface with the Shaker K+ channel. The fold of agitoxin is homologous to the previously determined folds of scorpion venom toxins. However, agitoxin 2 differs significantly from the other channel blockers in the specificity of its interactions. This study was thus focused on a precise characterization of the surface residues at the face of the protein interacting with the Shaker K+ channel. The rigid toxin molecule can be used to estimate dimensions of the potassium channel. Surface‐exposed residues, Arg24, Lys27, and Arg31 of the β‐sheet, have been identified from mutagenesis studies as functionally important for blocking the Shaker K+ channel. The sequential and spatial locations of Arg24 and Arg31 are not conserved among the homologous toxins. Knowledge on the details of the channel‐binding sites of agitoxin 2 formed a basis for site‐directed mutagenesis studies of the toxin and the K+ channel sequences. Observed interactions between mutated toxin and channel are being used to elucidate the channel structure and mechanisms of channel‐toxin interactions.Keywords
This publication has 40 references indexed in Scilit:
- Revealing the Architecture of a K + Channel Pore Through Mutant Cycles with a Peptide InhibitorScience, 1995
- Kaliotoxin (1-37) Shows Structural Differences With Related Potassium Channel BlockersBiochemistry, 1994
- Purification and Characterization of Three Inhibitors of Voltage-Dependent K+ Channels from Leiurus Quinquestriatus var. Hebraeus VenomBiochemistry, 1994
- Intimations of potassium channel structure from a complete functional map of the molecular surface of charybdotoxinBiochemistry, 1994
- Determination of the three-dimensional structure of iberiotoxin in solution by proton nuclear magnetic resonance spectroscopyBiochemistry, 1992
- Mechanism of iberiotoxin block of the large-conductance calcium-activated potassium channel from bovine aortic smooth muscleBiochemistry, 1992
- Design, synthesis, and functional expression of a gene for charybdotoxin, a peptide blocker of K+ channels.Proceedings of the National Academy of Sciences of the United States of America, 1991
- Determination of the subunit stoichiometry of a voltage-activated potassium channelNature, 1991
- Mechanism of charybdotoxin block of the high-conductance, Ca2+-activated K+ channel.The Journal of general physiology, 1988
- Charybdotoxin, a protein inhibitor of single Ca2+-activated K+ channels from mammalian skeletal muscleNature, 1985