Solution Structures of a Highly Insecticidal Recombinant Scorpion α-Toxin and a Mutant with Increased Activity,
- 1 March 1997
- journal article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 36 (9) , 2414-2424
- https://doi.org/10.1021/bi961497l
Abstract
The solution structure of a recombinant active alpha-neurotoxin from Leiurus quinquestriatus hebraeus, Lqh(alpha)IT, was determined by proton two-dimensional nuclear magnetic resonance spectroscopy (2D NMR). This toxin is the most insecticidal among scorpion alpha-neurotoxins and, therefore, serves as a model for clarifying the structural basis for their biological activity and selective toxicity. A set of 29 structures was generated without constraint violations exceeding 0.4 A. These structures had root mean square deviations of 0.49 and 1.00 A with respect to the average structure for backbone atoms and all heavy atoms, respectively. Similarly to other scorpion toxins, the structure of Lqh(alpha)IT consists of an alpha-helix, a three-strand antiparallel beta-sheet, three type I tight turns, a five-residue turn, and a hydrophobic patch that includes tyrosine and tryptophan rings in a "herringbone" arrangement. Positive phi angles were found for Ala50 and Asn11, suggesting their proximity to functionally important regions of the molecule. The sample exhibited conformational heterogeneity over a wide range of experimental conditions, and two conformations were observed for the majority of protein residues. The ratio between these conformations was temperature-dependent, and the rate of their interconversions was estimated to be on the order of 1-5 s(-1) at 308 K. The conformation of the polypeptide backbone of Lqh(alpha)IT is very similar to that of the most active antimammalian scorpion alpha-toxin, AaHII, from Androctonus australis Hector (60% amino acid sequence homology). Yet, several important differences were observed at the 5-residue turn comprising residues Lys8-Cys12, the C-terminal segment, and the mutual disposition of these two regions. 2D NMR studies of the R64H mutant, which is 3 times more toxic than the unmodified Lqh(alpha)IT, demonstrated the importance of the spatial orientation of the last residue side chain for toxicity of Lqh(alpha)IT.Keywords
This publication has 20 references indexed in Scilit:
- Binding of an α scorpion toxin to insect sodium channels is not dependent on membrane potentialFEBS Letters, 1993
- Structure of scorpion toxin variant-3 at 1·2 Å resolutionJournal of Molecular Biology, 1992
- Relationship between nuclear magnetic resonance chemical shift and protein secondary structureJournal of Molecular Biology, 1991
- Structure/activity relationships of scorpion α‐toxinsEuropean Journal of Biochemistry, 1989
- Determination of three‐dimensional structures of proteins from interproton distance data by hybrid distance geometry‐dynamical simulated annealing calculationsFEBS Letters, 1988
- Protein structures in solution by nuclear magnetic resonance and distance geometryJournal of Molecular Biology, 1987
- Stereospecific assignments of 1H‐nmr methyl lines and conformation of valyl residues in the lac repressor headpieceBiopolymers, 1985
- Primary structure of toxin IV of Leiurus quinquestriatus quinquestriatusFEBS Letters, 1985
- Pseudo-structures for the 20 common amino acids for use in studies of protein conformations by measurements of intramolecular proton-proton distance constraints with nuclear magnetic resonanceJournal of Molecular Biology, 1983
- α‐Scorpion neurotoxin derivatives suitable as potential markers of sodium channels. Preparation and characterizationInternational Journal of Peptide and Protein Research, 1983