As observed in Fig. 1, the selectivity of the CGTX-II, δ-AITX-Bcg1a and δ-AITX-Bcg1b toxins is highest

for Nav1.5 followed by 1.6 and 1.1 (Nav1.5 > 1.6 > 1.1). δ-AITX-Bcg1b KU-60019 was not shown to be potent and was consequently abandoned in our investigation. It is important to remind that δ-AITX-Bcg1b presents the single N16D substitution in relation to its isoform δ-AITX-Bcg1a (see Table 1). The latter shows a much higher potency among the assayed channels. However, CGTX-II also presents a D16 amino acid (see Table 1), but its potency and selectivity are close to the observed for δ-AITX-Bcg1a. In that case, it is clear that the N16 amino acid alone should not be considered as a key determinant of the potency or activity of sea anemone peptides. In the work by Oliveira et al. [23], the selectivity of ATX-II

(see its primary sequence in Table 1) was Nav1.1–1.2 > 1.5 > 1.4 > 1.6 > 1.3, selleck chemicals llc while its isoform AFT-II (with an extra Gly at N-terminus and a single K36A substitution, in relation to ATX-II) was selective as Nav1.4 > 1.5 > 1.6 > 1.3–1.1 > 1.2. The toxin BcIII (more alike to CGTX-II) was assayed in that work, showing a preferential activity on Nav1.5–1.1 > 1.4–1.6 > 1.2–1.3. More recently, these three peptides were assayed in Nav1.7 and all of them showed a smaller potency in that channel [34], such as for CGTX-II, δ-AITX-Bcg1a and δ-AITX-Bcg1b here presented. The compilation of those

data, together with a summary of the dose–response curves in the present study, is shown in Fig. 4. Contrary to AFT-II and BcIII, none Florfenicol of the toxins employed in this study showed some preference for binding to Nav1.4. Thus, it is clear that the selectivity of sea anemone type 1 toxins is variable, and consequently the surface of contact of each peptide should vary as well. Other authors tried to investigate this aspect [22]. They did a full alanine scanning of ATX-II (Av2) toxin, and found that some residues important for activity coincide, but many do not overlap with the contact surface of the structurally related peptide ApB [5], [10] and [31]. On the other hand, although differing only by N16D substitution, previous studies demonstrated that BgII (Asn) is much more potent than BgIII (Asp) (see Table 1) [6] and [28]. Consequently, this confirms that the role of each individual amino acid must be carefully examined for each toxin, and a single amino acid residue might not be as critical for binding on one isoform as for other. Very interestingly, our present data show that all the three toxins tested do not have a high preference for binding on Nav1.2 (the preferential target of ATX-II, one of the most potent sea anemone toxins). However, the supposed binding site (site 3) [8] of these type 1 sea anemone toxins in Nav1.1 is identical [23] and [30].