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PMID:15308644
| Citation |
Tedford, HW, Gilles, N, Ménez, A, Doering, CJ, Zamponi, GW and King, GF (2004) Scanning mutagenesis of omega-atracotoxin-Hv1a reveals a spatially restricted epitope that confers selective activity against insect calcium channels. J. Biol. Chem. 279:44133-40 |
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| Abstract |
We constructed a complete panel of alanine mutants of the insect-specific calcium channel blocker omega-atracotoxin-Hv1a. Lethality assays using these mutant toxins identified three spatially contiguous residues, Pro10, Asn27, and Arg35, that are critical for insecticidal activity against flies (Musca domestica) and crickets (Acheta domestica). Competitive binding assays using radiolabeled omega-atracotoxin-Hv1a and neuronal membranes prepared from the heads of American cockroaches (Periplaneta americana) confirmed the importance of these three residues for binding of the toxin to target calcium channels presumably expressed in the insect membranes. At concentrations up to 10 microm, omega-atracotoxin-Hv1a had no effect on heterologously expressed rat Cav2.1, Cav2.2, and Cav1.2 calcium channels, consistent with the previously reported insect selectivity of the toxin. 30 microm omega-atracotoxin-Hv1a inhibited rat Cav currents by 10-34%, depending on the channel subtype, and this low level of inhibition was essentially unchanged when Asn27 and Arg35, which appears to be critical for interaction of the toxin with insect Cav channels, were both mutated to alanine. We propose that the spatially contiguous epitope formed by Pro10, Asn27, and Arg35 confers specific binding to insect Cav channels and is largely responsible for the remarkable phyletic selectivity of omega-atracotoxin-Hv1a. This epitope provides a structural template for rational design of chemical insecticides that selectively target insect Cav channels. |
| Links |
PubMed Online version:10.1074/jbc.M404006200 |
| Keywords |
Amino Acid Substitution; Animals; Calcium Channel Blockers/pharmacology; Calcium Channels/immunology; Cockroaches; Gryllidae; Houseflies; Mutagenesis, Site-Directed; Protein Conformation; Rats; Spider Venoms/genetics; Spider Venoms/immunology; Spiders |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0019855: calcium channel inhibitor activity |
ECO:0000314: |
F |
Fig. 3B "Effect of ω-ACTX-Hv1a on rat HVA calcium channels...extent of current blockage for each channel subtype caused by either 30 μm ω-ACTX-Hv1a" |
complete | ||||
|
enables |
GO:0019855: calcium channel inhibitor activity |
ECO:0000314: direct assay evidence used in manual assertion |
F |
Seeded From UniProt |
complete | |||
See also
References
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