Comparison of the structure and dynamics of the antibiotic peptide polymyxin B and the inactive nonapeptide in aqueous trifluoroethanol by NMR spectroscopy.

The structure and dynamics of polymyxin B (PxB), an N-acylated cyclic decapeptide that displays antimicrobial activity against Gram-negative bacteria, is characterized by NMR and compared to results for the inactive nonapeptide, which is missing the N-terminal amino acid along with the attached acyl chain. Aqueous trifluoroethanol (TFE) was chosen as the solvent since the overall structure of PxB in TFE is similar to the structure when bound to vesicles. No differences were observed between the two peptides for (1)H H(alpha) chemical shifts or patterns of cross peaks in NOESY spectra, indicating that the overall structures are quite similar. The sign and intensity of NOESY spectra obtained at different temperatures were used to assess the relative mobility of the peptides. For both peptides, differential mobility is observed in different parts of the molecule, with greater mobility observed for the linear portion than the ring and faster motion seen for the side chains than the peptide backbone. However, all motion is faster in the nonapeptide, indicating that the presence of the N-terminal acyl chain restricts the mobility of PxB compared to the nonapeptide, which lacks this structural feature. For both peptides, differential mobility is also observed within the cyclic portion of the peptide. This supports a proposed model whereby the more rigid residues serve as pivot points, allowing the ring conformation to change in response to different binding partners. However, conformational flexibility within the cyclic ring is not sufficient for antimicrobial activity since both the active and inactive peptides exhibit the same flexibility. The N-terminal acyl chain on PxB, which is essential for activity, exhibits rapid, independent motion, and this flexibility may facilitate penetration of the outer membrane.