X-ray Structure and Mutagenesis of the Scorpion Depressant Toxin LqhIT2 Reveals Key Determinants Crucial for Activity and Anti-Insect Selectivity

Journal of Molecular Biology

Volumn 366, Issue 2, 2007, Pages 586-601

  Karbat, Izhar ^a   Turkov, Michael ^a   Cohen, Lior ^a   Kahn, Roy ^a   Gordon, Dalia ^a   Gurevitz, Michael ^a   Frolow, Felix ^a  

^a TEL AVIV UNIVERSITY   (Israel)

Author keywords

anti insect selectivity; bioactive surface; scorpion depressant toxin; X ray structure

Indexed keywords

ALANINE; ASPARAGINE; CYSTEINE; GLUTAMIC ACID; LQHIT2 TOXIN; SCORPION VENOM; TRYPTOPHAN; TYROSINE; UNCLASSIFIED DRUG; VOLTAGE GATED SODIUM CHANNEL;

AMINO ACID SUBSTITUTION; ARTICLE; BINDING AFFINITY; CONFORMATIONAL TRANSITION; HYDROPHOBICITY; INSECT; LEIURUS QUINQUESTRIATUS HEBRAEUS; MOLECULAR MECHANICS; MOLECULE; MUTAGENESIS; NONHUMAN; PHARMACOPHORE; PRIORITY JOURNAL; SCORPION; VOLTAGE SENSOR TRAPPING; X RAY;

HEXAPODA; LEIURUS QUINQUESTRIATUS HEBRAEUS; SCORPIONES;

EID: 33846396124     PISSN: 00222836     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.jmb.2006.10.085     Document Type: Article

References (44)

1. Lester D., Lazarovici P., Pelhate M., and Zlotkin E. Purification, characterization and action of two insect toxins from the venom of the scorpion buthotus judaicus. Biochim. Biophys. Acta 701 (1982) 370-381
2. Zlotkin E., Eitan M., Bindokas V.P., Adams M.E., Moyer M., Burkhart W., and Fowler E. Functional duality and structural uniqueness of depressant insect-selective neurotoxins. Biochemistry 30 (1991) 4814-4821
3. Strugatsky D., Zilberberg N., Stankiewicz M., Ilan N., Turkov M., Cohen L., et al. Genetic polymorphism and expression of a highly potent scorpion depressant toxin enable refinement of the effects on insect Na channels and illuminate the key role of Asn-58. Biochemistry 44 (2005) 9179-9187
4. Gordon D., Jover E., Couraud F., and Zlotkin E. The binding of an insect selective neurotoxin (AaIT) from scorpion venom to locust synaptosomal membranes. Biochim. Biophys. Acta 778 (1984) 349-358
5. Gordon D., Moskowitz H., Eitan M., Warner C., Catterall W.A., and Zlotkin E. Localization of receptor sites for insect-selective toxins on sodium channels by site-directed antibodies. Biochemistry 31 (1992) 7622-7628
6. Gordon D., Savarin P., Gurevitz M., and Zinn-Justin S. Functional anatomy of scorpion toxins affecting sodium channels. J. Toxicol. Toxin Rev. 17 (1998) 131-158
7. Herrmann R., Moskowitz H., Zlotkin E., and Hammock B.D. Positive cooperativity among insecticidal scorpion neurotoxins. Toxicon 33 (1995) 1099-1102
8. Ben Khalifa R., Stankiewicz M., Pelhate M., Serrano-Hernandez S.E., Possani L.D., Hinkel H., and Mebs D. Action of babycurus-toxin 1 from the east African scorpion Babycurus centrurimorphus on the isolated cockroach giant axon. Toxicon 35 (1997) 1069-1080
9. Cestele S., Kopeyan C., Oughideni R., Mansuelle P., Granier C., and Rochat H. Biochemical and pharmacological characterization of a depressant insect toxin from the venom of the scorpion Buthacus arenicola. Eur. J. Biochem. 243 (1997) 93-99
10. Ali S.A., Storva S., Grossmann J.G., Abbasi A., and Voelter W. Purification, characterization, and primary structure of four depressant insect-selective neurotoxin analogs from scorpion (Buthus sindicus) venom. Arch. Biochem. Biophys. 391 (2001) 197-206
11. Borchani L., Stankiewicz M., Kopeyan C., Mansuelle P., Kharrat R., Cestele S., et al. Purification, structure and activity of three insect toxins from Buthus occitanus tunetanus venom. Toxicon 35 (1997) 365-382
12. Kopeyan C., Mansuelle P., Sampieri F., Brando T., Bahraoui E.M., Rochat H., and Granier C. Primary structure of scorpion anti-insect toxins isolated from the venom of Leiurus quinquestriatus. FEBS Letters 261 (1990) 423-426
13. Zlotkin E., Kadouri D., Gordon D., Pelhate M., Martin M.-F., and Rochat H. An excitatory and a depressant insect toxin from scorpion venom both affect sodium conductance and possess a common binding site. Arch. Biochem. Biophys. 240 (1985) 877-887
14. Nakagawa Y., Lee Y.M., Lehmberg E., Herrmann R., Moskowitz H., Jones A.D., and Hammock B.D. Anti-insect toxin (AaIT5) from Androctonus australis. Eur. J. Biochem. 246 (1997) 496-501
15. Mejri T., Borchani L., Srairi-Abid N., Benkhalifa R., Cestele S., Regaya I., et al. BotIT6: a potent depressant insect toxin from Buthus occitanus tunetanus venom. Toxicon 41 (2003) 163-171
16. Possani L.D., Becerril B., Delepierre M., and Tytgat J. Scorpion toxins specific for Na+-channels. Eur. J. Biochem. 264 (1999) 287-300
17. + channels and related peptides: biodiversity, structure-function relationships and evolution. Toxicon 46 (2005) 831-844
18. Pintar A., Possani L.D., and Delepierre M. Solution structure of toxin 2 from Centruroides noxius Hoffmann, a β-scorpion neurotoxin acting on sodium channels. J. Mol. Biol. 287 (1999) 359-367
19. Froy O., and Gurevitz M. Membrane potential modulators: a thread of scarlet from plants to humans. FASEB J. 12 (1998) 1793-1796
20. Oren D.A., Froy O., Amit E., Kleinberger-Doron N., Gurevitz M., and Shaanan B. An excitatory scorpion toxin with a distinctive feature: an additional alpha helix at the C terminus and its implications for interaction with insect sodium channels. Structure 6 (1998) 1095-1103
21. Cohen L., Karbat I., Gilles N., Ilan N., Benveniste M., Gordon D., and Gurevitz M. Common features in the functional surface of scorpion beta-toxins and elements that confer specificity for insect and mammalian voltage-gated sodium channels. J. Biol. Chem. 280 (2005) 5045-5053
22. + channels. J. Biol. Chem. 279 (2004) 8206-8211
23. Cestele S., Qu Y., Rogers J.C., Rochat H., Scheuer T., and Catterall W.A. Voltage sensor-trapping: enhanced activation of sodium channels by beta-scorpion toxin bound to the S3-S4 loop in domain II. Neuron 21 (1998) 919-931
24. + channels. FASEB J. 18 (2004) 683-689
25. Zhao B., Carson M., Ealick S.E., and Bugg C.E. Structure of scorpion toxin variant-3 at 1.2 Å resolution. J. Mol. Biol. 227 (1992) 239-252
26. + channels. Acta Crystallog. sect. D 59 (2003) 405-415
27. Shatsky M., Nussinov R., and Wolfson H.J. A method for simultaneous alignment of multiple protein structures. Proteins: Struct. Funct. Genet. 56 (2004) 143-156
28. Derewenda Z.S., Lee L., and Derewenda U. The occurrence of C-H...O hydrogen bonds in proteins. J. Mol. Biol. 252 (1995) 248-262
29. Hassani O., Mansuelle P., Cestele S., Bourdeaux M., Rochat H., and Sampieri F. Role of lysine and tryptophan residues in the biological activity of toxin VII (Ts gamma) from the scorpion Tityus serrulatus. Eur. J. Biochem. 260 (1999) 76-86
30. Bogan A.A., and Thorn K.S. Anatomy of hot spots in protein interfaces. J. Mol. Biol. 280 (1998) 1-9
31. Tong X., Yao J., He F., Chen X., Zheng X., Xie C., et al. NMR solution structure of BmK-betaIT, an excitatory scorpion beta-toxin without a 'hot spot' at the relevant position. Biochem. Biophys. Res. Commun. 349 (2006) 890-899
32. Turkov M., Rashi S., Zilberberg N., Gordon D., Ben Khalifa R., Stankiewicz M., et al. In vitro folding and functional analysis of an anti-insect selective scorpion depressant neurotoxin produced in Escherichia coli. Protein Express. Purific. 10 (1997) 123-131
33. Reed L., and Muench H. A simple method of estimating fifty-percent endpoint. Am. J. Hyg. 27 (1938) 493-497
34. + current in Xenopus oocytes. J. Neurosci. 12 (1992) 3743-3752
35. Wallner M., Weigl L., Meera P., and Lotan I. Modulation of the skeletal muscle sodium channel alpha-subunit by the beta 1-subunit. FEBS Letters 336 (1993) 535-539
36. Shichor I., Zlotkin E., Ilan N., Chikashvili D., Stuhmer W., Gordon D., and Lotan I. Domain 2 of Drosophila para voltage-gated sodium channel confers insect properties to a rat brain channel. J. Neurosci. 22 (2002) 4364-4371
37. Teng T.Y. Mounting of crystals for macromolecular crystallography in a freestanding thin-film. J. Appl. Crystallog. 23 (1990) 387-391
38. Cosier J., and Glazer A.M. A nitrogen-gas-stream cryostat for general X-ray-diffraction studies. J. Appl. Crystallog. 19 (1986) 105-107
39. Otwinowski Z., and Minor W. Processing of X-ray diffraction data collected in oscillation mode. Methods Enzymol. 276 (1997) 307-326
40. Bailey S. The CCP4 Suite - programs for protein crystallography. Acta Crystallog. sect. D 50 (1994) 760-763
41. Vagin A., and Teplyakov A. MOLREP: an automated program for molecular replacement. J. Appl. Crystallog. 30 (1997) 1022-1025
42. Murshudov G.N., Vagin A.A., and Dodson E.J. Refinement of macromolecular structures by the maximum-likelihood method. Acta Crystallog. sect. D 53 (1997) 240-255
43. Morris R.J., Perrakis A., and Lamzin V.S. ARP/wARP and automatic interpretation of protein electron density maps. Methods Enzymol. 374 (2003) 229-244
44. Emsley P., and Cowtan K. Coot: model-building tools for molecular graphics. Acta Crystallog. sec. D 60 (2004) 2126-2132