Conformational changes of alamethicin induced by solvent and temperature. A 13C-NMR and circular-dichroism study.
13C nuclear magnetic resonance (NMR) and circular dichroism (CD) have been used for studies on the conformation of alamethicin. The 13C NMR spectrum is assigned with the aid of signals of synthetic partial sequences and selective proton decoupling. The solvent and temperature-dependence of the 13C NMR spectra, T1 measurements and the use of lanthanide-shift reagents allow the differentiation between the amino acids belonging to a rigid alpha-helical portion of the alamethicin sequence and those belonging to a more flexible part. The 13C NMR results are in agreement with results obtained from extended solvent and temperature-dependent CD studies which indicate a highly stabilized nonpolar and intrachenar alpha-helical part. The concentration-dependence of the CD spectrum of alamethicin in a nematic phase revealed aggregation phenomena which might simulate those observed in natural and synthetic membranes. After dissolving alamethicin in aqueous alcohol there is a time-dependence of the ellipticity of the Cotton effects showing a sort of memory effect on the mode of dissolution. Four different conformations can be characterized by CD spectra depending on the solvent and concentration. A model illustrating the dynamic conformations and aggregation phenomena within a membrane is proposed.
D000408:Alamethicin; D000900:Anti-Bacterial Agents; D001665:Binding Sites; D002247:Carbon Isotopes; D002942:Circular Dichroism; D005583:Fourier Analysis; D009682:Magnetic Resonance Spectroscopy; D011485:Protein Binding; D011487:Protein Conformation; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature
D000900:Anti-Bacterial Agents; D002247:Carbon Isotopes; D012997:Solvents; D000408:Alamethicin