EXPERIMENTAL MOLECULAR-DYNAMICS OF AN ALANINE-BASED HELICAL PEPTIDE DETERMINED BY SPIN-LABEL ELECTRON-SPIN-RESONANCE

The alanine-based 3K(I) peptide is reported to be very helical in aqueous solution. We have prepared a series of six nitroxide spin labeled analogs of the 3K(I) sequence and measured the variable-temperature ESR spectra for each in order to reveal the position-dependent peptide dynamics. From analysis of these local dynamics under helix-forming conditions at 1-degrees-C, we find that the helix termini show greater local dynamics than the peptide center. Further, the C-terminus is more mobile than the N-terminus. Even in the helix-promoting solvent trifluoroethanol, the results indicate that there is still substantially greater dynamics at the helix termini than at the peptide center. The unfolded state is also investigated, and we find that the peptide unfolded by guanidine hydrochloride is somewhat different than that found for high-temperature aqueous solution. Recently it was suggested that short 16-mer peptides may adopt a 3(10)-helix structure instead of the expected alpha-helix. The data presented here at 1-degrees-C show that there is sufficient disorder with in the peptide to accommodate the 3(10) structure. Also calculated are the backbone torsional fluctuations, and the results compare well to those from computer molecular dynamics studies. A proposal is outlined that explains how the enhanced dynamics found at the C-terminus results from the exposure of the helix hydrogen bonds to aqueous solvent in this region of the peptide.