Synthetic peptide fragments as probes for structure determination of potassium ion-channel proteins.
Potassium channels are a diverse class of transmembrane proteins that are responsible for diffusion of potassium ion across cell membranes. The lack of large quantities of these proteins from natural sources, is a major hindrance in their structural characterization using biophysical techniques. Synthetic peptide fragments corresponding to functionally important domains of these proteins provide an attractive approach towards characterizing the structural organization of these ion-channels. Conformational properties of peptides from three different potassium channels (Shaker, ROMK1 and minK) have been characterized in aqueous media, organic solvents and in phospholipid membranes. Techniques used for these studies include FTIR, CD and 2D-NMR spectroscopy. FTIR spectroscopy has been a particularly valuable tool for characterizing the folding of the ion-channel peptides in phospholipid membranes; the three different types of potassium channels all share a common transmembrane folding pattern that is composed of a predominantly alpha-helical structure. There is no evidence to suggest the presence of any significant beta-sheet structure. These results are in excellent agreement with the crystal structure of a bacterial potassium channel (Doyle, D. A. et al. (1998) Science 280:69 77), and suggest that all potassium channel proteins may share a common folding motif where the ion-channel structure is constructed entirely from alpha-helices.
Haris
P I
PI
Department of Biological Sciences, De Montfort University, Leicester, United Kingdom.
D001426:Bacterial Proteins; D010446:Peptide Fragments / Q000138:chemical synthesis*; D015221:Potassium Channels / Q000737:chemistry*; D024661:Potassium Channels, Inwardly Rectifying; D024642:Potassium Channels, Voltage-Gated; D011487:Protein Conformation; D051637:Shaker Superfamily of Potassium Channels; D017550:Spectroscopy, Fourier Transform Infrared
D001426:Bacterial Proteins; D010446:Peptide Fragments; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D024642:Potassium Channels, Voltage-Gated; D051637:Shaker Superfamily of Potassium Channels; C059983:potassium channel protein I(sk); C107790:prokaryotic potassium channel