The secondary structure of the toxin ATX Ia from Anemonia sulcata in aqueous solution determined on the basis of complete sequence-specific 1H-NMR assignments.
The toxin preparations ATX I, ATX Ia and ATX Ib from Anemonia sulcata were investigated by proton nuclear magnetic resonance (NMR). High-resolution phase-sensitive two-dimensional NMR experiments were used to monitor the separation by high-performance liquid chromatography of the two isoproteins ATX Ia and ATX Ib. For ATX Ia complete sequence-specific resonance assignments were obtained and the secondary structure was determined. To obtain the NMR assignments we used a variant of the sequential assignment technique which relied extensively on cross-peak fine-structure analysis in phase-sensitive spectra, using spectrum simulations based on density matrix calculations with the program SPHINX. These procedures, which resulted in extensive amino acid spin system identifications prior to the sequential assignments, should be generally applicable for small proteins with relatively narrow 1H-NMR lines. The secondary structure of ATX I includes a beta sheet consisting of four strands. No evidence was found for the presence of regular helical segments. The four beta strands are connected by two extended loops and a tight turn, for which further characterization has to await the complete determination of the three-dimensional structure.
D000595:Amino Acid Sequence; D003064:Cnidarian Venoms; D006860:Hydrogen Bonding; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010455:Peptides; D011487:Protein Conformation; D012996:Solutions
D003064:Cnidarian Venoms; D010455:Peptides; D012996:Solutions