8.B.1 The Long (4C-C) Scorpion Toxin (L-ST) Superfamily The NaC- L-ST superfamily contains a large number of scorpion-derived peptide toxins. These are tabulated below with descriptions. They include the well-characterized scorpion α- and β-toxins that act on tetrodotoxin-inhibitable, voltage-gated Na⁺ channels (TC #1.A.1.10). While the α-toxins (e.g., from Buthinea venom) prolong the Na⁺-inactivation phase of the activated action potential-causing channels, thereby blocking neuronal transmission, the β-toxins (e.g., from Centrurinae sculpturatus venom) affect the Na⁺-activation phase. These toxins are derived from a variety of scorpions. They can affect both insect and mammalian Na⁺ channel activities (Tian et al., 2007). In many cases the details of their toxic actions are known. β-scorpion toxin, for example, targets neurotoxin receptor site 4 in Na⁺ channels and induces a negative shift in the voltage dependence of activation through a voltage sensor-trapping mechanism (Cestèle et al., 2006). A single organism may produce many of these toxins, some closely related, others more distantly related. This superfamily includes hundreds of sequenced members as revealed by PSI-BLAST searches with six iterations, some of which are reported to be non-toxic. They are usually characterized by four disulfide bridges, but some have three or five. Among scorpion species, the Buthidae produce the most deadly and painful venoms. A pain-inducing α-toxin (CvIV4) was isolated from the venom of Centruroides vittatus and tested on five Na⁺ channel isoforms (Rowe et al. 2011). CvIV4 slowed the fast inactivation of Na(v)1.7 (TC# 1.A.1.10.5), a Na⁺ channel expressed in peripheral pain-pathway neurons (nociceptors), but did not affect the Na(v)1.8-based sodium currents of these neurons (TC# 1.A.1.10.6). CvIV4 also slowed the fast inactivation of Na(v)1.2, Na(v)1.3 and Na(v)1.4. The effects of CvIV4 are similar to Old World α-toxins that target Na(v)1.7 (AahII, BmK MI, LqhIII, OD1), but the primary sequence of CvIV4 is not similar to these toxins. Mutant Na(v)1.7 channels (D1586A and E1589Q, DIV S3-S4 linker) reduced but did not abolish the effects of CvIV4.
This family belongs to the Defensin Superfamily.
References:
Cestèle, S., T. Scheuer, M. Mantegazza, H. Rochat, and W.A. Catterall. (2001). Neutralization of gating charges in domain II of the sodium channel alpha subunit enhances voltage-sensor trapping by a β-scorpion toxin. J Gen Physiol 118: 291-302.
Cestèle, S., Y. Qu, J.C. Rogers, H. Rochat, T. Scheuer, and W.A. Catterall. (1998). Voltage sensor-trapping: enhanced activation of sodium channels by β-scorpion toxin bound to the S3-S4 loop in domain II. Neuron. 21: 919-931.
Cestèle, S., Yarov-Yarovoy, V., Qu, Y., Sampieri, F., Scheuer, T., and Catterall, W.A. (2006). Structure and function of the voltage sensor of sodium channels probed by a β-scorpion toxin. J. Biol. Chem. 281: 21332-21344.
Cohen, L., Y. Moran, A. Sharon, D. Segal, D. Gordon, and M. Gurevitz. (2009). Drosomycin, an innate immunity peptide of Drosophila melanogaster, interacts with the fly voltage-gated sodium channel. J. Biol. Chem. 284: 23558-23563.
del Río-Portilla, F., E. Hernández-Marín, G. Pimienta, F.V. Coronas, F.Z. Zamudio, R.C. Rodríguez de la Vega, E. Wanke, and L.D. Possani. (2004). NMR solution structure of Cn12, a novel peptide from the Mexican scorpion Centruroides noxius with a typical β-toxin sequence but with α-like physiological activity. Eur J Biochem 271: 2504-2516.
Fontecilla-Camps, J.C., Habersetzer-Rochat, C., and Rochat, H. (1988). Orthorhombic crystals and three-dimensional structure of the potent toxin II from the scorpion Androctonus australis Hector. Proc. Natl. Acad. Sci. USA 85: 7443-7447.
Guan, R.J., Xiang, Y., He, X.L., Wang, C.G., Wang, M., Zhang, Y., Sundberg, E.J., and Wang, D.C. (2004). Structural mechanism governing cis and trans isomeric states and an intramolecular switch for cis/trans isomerization of a non-proline peptide bond observed in crystal structures of scorpion toxins. J. Mol. Biol. 341: 1189-1204.
Ma, Z., J. Kong, D. Gordon, M. Gurevitz, and R.G. Kallen. (2013). Direct Evidence that Scorpion α-Toxins (Site-3) Modulate Sodium Channel Inactivation by Hindrance of Voltage-Sensor Movements. PLoS One 8: e77758.
Mantegazza, M. and S. Cestèle. (2005). Beta-scorpion toxin effects suggest electrostatic interactions in domain II of voltage-dependent sodium channels. J. Physiol. 568: 13-30.
Possani, L.D., Becerril, B., Delepierre, M., and Tytgat, J. (1999). Scorpion toxins specific for Na⁺-channels. Eur. J. Biochem. 264: 287-300.
Rowe, A.H., Y. Xiao, J. Scales, K.D. Linse, M.P. Rowe, T.R. Cummins, and H.H. Zakon. (2011). Isolation and characterization of CvIV4: a pain inducing α-scorpion toxin. PLoS One 6: e23520.
Tian, C., Y. Yuan, and S. Zhu. (2008). Positively selected sites of scorpion depressant toxins: possible roles in toxin functional divergence. Toxicon 51(4): 555-562.
Zhu, S., B. Gao, M. Deng, Y. Yuan, L. Luo, S. Peigneur, Y. Xiao, S. Liang, and J. Tytgat. (2010). Drosotoxin, a selective inhibitor of tetrodotoxin-resistant sodium channels. Biochem Pharmacol 80: 1296-1302.
Zuo, X.P. and Ji, Y.H. (2004). Molecular mechanism of scorpion neurotoxins acting on sodium channels: insight into their diverse selectivity. Mol. Neurobiol. 30: 265-278.
Examples:
TC#	Name	Organismal Type	Example
8.B.1.1.1	*The α-neurotoxin BmK-MI precursor/x-ray structure known to 1.4 Å (Guan et al., 2004) (α-subfamily)*	Scorpions	*BmK-MI of Mesobuthus (Buthus) martensii* (84 aas; P45697)**

8.B.1.1.10	U3m-buthitoxin-Hj1a, partial, of 83 aas.		Hj1a of Hottentotta judaicus

8.B.1.1.11	Toxin TdNa9 of 85 aas and 1 TMS. This toxin binds, in vitro, to sodium channels and inhibits the inactivation of the activated channels.		*TdNa9 of Tityus discrepans* (Venezuelan scorpion)**

8.B.1.1.12	Galiomycin of 68 aas and 1 N-terminal TMS. It is an antifungal agent.		Gali of Helicoverpa zea (Corn earworm moth) (Heliothis zea)

8.B.1.1.13	Scorpion toxin, Cn12 of 67 aas and four disulfide bridges. he NMR structure has been determined. It has a beta-toxin sequence but alpha-like physiological activity (del Río-Portilla et al. 2004). It binds voltage-independently at site-3 of sodium channels (Nav) to inhibit the inactivation of the activated channels, thereby blocking neuronal transmission.		*Cn12 of Centruroides noxius* (Mexican scorpion)**

8.B.1.1.2	Markatoxin-III (MkTx-III) precursor	Scorpions	*MkTx-III of Mesobuthus (Buthus) martensii* (85 aas; P59853)**

8.B.1.1.3	Non-toxic immunogenic venom protein, NTxp, precursor	Scorpions	*NTxp of Tityus serrulatus* (84 aas; O77463)**

8.B.1.1.4	Depressant anti-insect-specific toxin 2 precursor, LqqIT2	Scorpions	*LqqIT2 of Leiurus quinquestriatus* (82 aas; P19855)**

8.B.1.1.5	Excitatory insect-selective toxin 1 precursor, BmKIT1 (binds voltage independently to Na⁺ channels, shifting the voltage of activation to more negative potentials)	Scorpions	*BmKIT1 of Buthus martensii* (88 aas; O61668)**

8.B.1.1.6	Pain-inducing α-toxin of 97 aas, CvIV4 (Rowe et al. 2011). CvIV4 slowed the fast inactivation of Na(v)1.7, a Na⁺ channel expressed in peripheral pain-pathway neurons (nociceptors), but did not affect the Na(v)1.8-based sodium currents of these neurons. CvIV4 also slowed the fast inactivation of Na(v)1.2, Na(v)1.3 and Na(v)1.4. The effects of CvIV4 are similar to Old World α-toxins that target Na(v)1.7 (AahII, BmK MI, LqhIII, OD1).	Scorpions	CvIV4 of Centruroides vittatus

8.B.1.1.7	AahII; neurotoxin 2 of 85 aas. Scorpion α-toxins bind voltage-independently at site-3 of sodium channels (Nav) and inhibit the inactivation of the activated channels, thereby blocking neuronal transmission. This toxin is active against mammals.	Animals	*AahII of Androctonus australis* (Sahara scorpion)**

8.B.1.1.8	α-Neurotoxin of 67 aas from the yellow scorpion, LqhIII. Binds voltage-independently at site-3 of sodium channels and inhibits the inactivation of the activated channels, modulating inactivation by hindering voltage-sensor movement, thereby blocking neuronal transmission (Ma et al. 2013). Dissociation is voltage-dependent. This alpha-like toxin is highly toxic to insects and competes with LqhaIT on binding to insect sodium channels. Differs from classical anti-mammalian alpha-toxins as it inhibits sodium channel inactivation in cell bodies of hippocampus brain neurons, on which the anti-mammalian Lqh2 is inactive, and is unable to affect Nav1.2 in the rat brain, on which Lqh2 is highly active (Rowe et al. 2011).	Animals (scorpions)	LqhIII of Leiurus quinquestriatus hebraeus

8.B.1.1.9	Drosomycin, an antimicrobial antifungal peptide of 70 aas and 1 N-terminal TMS. Targets tetrodttoxin resistant Na⁺ channels (Zhu et al. 2010) including the Drosophila Na⁺ channel (Cohen et al. 2009).	Animals	Drosomycin of Drosophila melanogaster

Examples:
TC#	Name	Organismal Type	Example
8.B.1.2.1	Anti-crustacean-specific toxin 1 (β-subfamily)	Scorpions	*Anti-crustacean-specific toxin 1 of Centruroides limpidus* (66 aas; P45667)**

8.B.1.2.2	Beta-anti-mammalian scorpian toxin, Css4 of 87 aas and 1 N-terminal TMS. Beta toxins bind voltage-independently at site-4 of sodium channels (Nav) and shift the voltage of activation toward more negative potentials, thereby affecting sodium channel activation and promoting spontaneous and repetitive firing (Cestèle et al. 2001). This toxin is active only on Na⁺ channels in mammals (Cestèle et al. 1998). Binding results from electrostatic interactions in domain II of the Na⁺ channels (Mantegazza and Cestèle 2005).		*Css4 of Centruroides suffusus* (Durango bark scorpion)**

Examples:
TC#	Name	Organismal Type	Example
8.B.1.3.1	Uncharacterized protein of 100 aas and 1 N-terminal TMS.		UP of Setaria viridis

Examples:
TC#	Name	Organismal Type	Example
8.B.1.4.1	Conotoxin Mr22.1 of 90 aas and 1 N-terminal TMS. This protein aligns with 9.B.1.1.9.		Conotoxin of Conus marmoreus

8.B.1.4.2	Conotoxin precursor superfamily E, partial, of 86 aas and 1 N-terminal TMS.		Conotoxin of Conus ermineus

8.B.1.4.3	Conotoxin Vc22.1 of 91 aas and 1 N-terminal TMS.		*C-Vc22 of Conus victoriae* (Queen Victoria cone)**

8.B.1.4.4	Conopeptide im005 of 83 aas and 1 N-terminal TMS.		C-im005 of Conus imperialis