FIGURE 1.

Isolation of the active peptide from the venom of U. grebelnyi. Fractions marked with a black box exercised an inhibitory activity on the ASIC3 channel in all separation steps. A, the first separation stage of the crude peptide fraction by size exclusion chromatography on a TSK-2000SW (7.5 × 600 mm) column in 10% acetonitrile solution with 0.1% trifluoroacetic acid and a flow rate of 0.5 ml/min. B, second separation step on a reverse-phase column Jupiter C₅ column (4.6 × 150 mm) in 0.1% trifluoroacetic acid with a flow rate of 1 ml/min using a linear gradient of acetonitrile concentration. C, final separation step on a Vydac C₁₈ column (250 × 4.6 mm) using a linear gradient of acetonitrile concentration in 0.1% trifluoroacetic acid with a flow rate 1 ml/min.