Structural characterization of a paramagnetic metal-ion-assembled three-stranded alpha-helical coiled coil.

A helical peptide designed to present an all-leucine core upon folding has been shown to exhibit concentration-dependent helicity and to exist as an ill-defined equilibrium population of oligomers. In marked contrast, an identical peptide covalently modified with a 2,2'-bipyridyl group at the N terminus forms a stable three-stranded parallel coiled coil in the presence of transition metal ions. We have employed paramagnetic Ni(2+) and Co(2+) ions to stabilize the trimeric assembly and to exploit their shift and relaxation properties in NMR structural studies. We find that metal-ion binding and helix-bundle folding are tightly coupled. Surprisingly, the three-helix bundle exhibits a dynamic N-terminal region, and a well-structured C-terminal half. The spectra indicate the presence of a dual conformation for the bundle extending from the N terminus to residue 12. The structure of the two isomeric forms has been ascertained from interpretation of NOEs in the Ni(II) complex and (1)H pseudocontact shifts in the Co(II) complex. Two different facial isomers with distinct susceptibility tensors were identified. The bulky leucine side chain at position 3 in the peptide chain appears to play a role in the conformational variation at the N terminus.