The 1.1 Å Resolution crystal structure of [Tyr¹⁵]EpI, a novel α- Conotoxin from Conus episcopatus, solved by direct methods

Conotoxins are valuable probes of receptors and ion channels because of their small size and highly selective activity. α-Conotoxin EpI, a 16- residue peptide from the mollusk-hunting Conus episcopatus, has the amino acid sequence GCCSDPRCNMNNPDY(SO₃H)C-NH₂ and appears to be an extremely potent and selective inhibitor of the α3β2 and α3β4 neuronal subtypes of the nicotinic acetylcholine receptor (nAChR). The desulfated form of EpI ([Tyr¹⁵]EpI) has a potency and selectivity for the nAChR receptor similar to those of EpI. Here we describe the crystal structure of [Tyr¹⁵]EpI solved at a resolution of 1.1 Å using SnB. The asymmetric unit has a total of 284 non-hydrogen atoms, making this one of the largest structures solved de novo by direct methods. The [Tyr¹⁵]EpI structure brings to six the number of α-conotoxin structures that have been determined to date. Four of these, [Tyr¹⁵]EpI, PnIA, PnIB, and MII, have an α4/7 cysteine framework and are selective for the neuronal subtype of the nAChR. The structure of [Tyr¹⁵]EpI has the same backbone fold as the other α4/7-conotoxin structures, supporting the notion that this conotoxin cysteine framework and spacing give rise to a conserved fold. The surface charge distribution of [Tyr¹⁵]EpI is similar to that of PnIA and PnIB but is likely to be different from that of MII, suggesting that [Tyr¹⁵]EpI and MII may have different binding modes for the same receptor subtype.