THE NMR SOLUTION STRUCTURE OF THE PHEROMONE ER-2 FROM THE CILIATED PROTOZOAN EUPLOTES-RAIKOVI

The NMR structure of the pheromone Er-2 from the ciliated protozoan Eu plotes raikovi has been determined in aqueous solution. The structure of this 40-residue protein was calculated with the distance geometry p rogram DIANA from 621 distance constraints and 89 dihedral angle const raints; the program OPAL was employed for the energy minimization. For a group of 20 conformers used to characterize the solution structure, the average pairwise RMS deviation from the mean structure calculated for the backbone heavy atoms N, C-alpha, and C' of residues 3-37 was 0.31 Angstrom. The molecular architecture is dominated by an up-down-u p bundle of 3 short helices of residues 5-11, 14-20, and 23-33, which is similar to the structures of the homologous pheromones Er-1 and Er- 10. Novel structural features include a well-defined N-cap on the firs t helix, a 1-residue deletion in the second helix resulting in the for mation of a 3(10)-helix rather than an alpha-helix as found in Er-1 an d Er-10, and the simultaneous presence of 2 different conformations fo r the C-terminal tetrapeptide segment, i.e., a major conformation with the Leu 39-Pro 40 peptide bond in the trans form and a minor conforma tion with this peptide bond in the cis form.