Conformation of gramicidin A channel in phospholipid vesicles: a 13C and 19F nuclear magnetic resonance study.
We have determined the conformation of the channel-forming polypeptide antibiotic gramicidin A in phosphatidylcholine vesicles by using 13C and 19F NMR spectroscopy. The models previously proposed for the conformation of the dimer channel differ in the surface localization of the NH2 and COOH termini. We have incorporated specific 13C and 19F nuclei at both the NH2, and COOH termini of gramicidin and have used 13C and 19F chemical shifts and spin lattice relaxation time measurements to determine the accessibility of these labels to three paramagnetic NMR probes--two in aqueous solution and one attached to the phosphatidylcholine fatty acid chain9 all of our results indicate that the COOH terminus of gramicidin in the channel is located near the surface of the membrane and the NH2 terminus is buried deep within the lipid bilayer. These findings strongly favor an NH2-terminal to NH2-terminal helical dimer as the major conformation for the gramicidin channel in phosphatidylcholine vesicles.
D002247:Carbon Isotopes; D005461:Fluorine; D006096:Gramicidin; D007473:Ion Channels / Q000187:drug effects; D008051:Lipid Bilayers; D009682:Magnetic Resonance Spectroscopy; D008345:Manganese / Q000494:pharmacology; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D011487:Protein Conformation; D013113:Spin Labels; D013932:Thulium / Q000494:pharmacology
D002247:Carbon Isotopes; D007473:Ion Channels; D008051:Lipid Bilayers; D008563:Membrane Lipids; D010713:Phosphatidylcholines; D013113:Spin Labels; D006096:Gramicidin; D005461:Fluorine; D008345:Manganese; D013932:Thulium