title: NMR structure of C-terminally tagged gramicidin channels.
abstract: A biotin group was covalently attached to the C terminus of gramicidin A (gA) through a linker arm comprising a glycine residue with either one (gAXB) or two caproyl groups (gAXXB). High-resolution two-dimensional NMR spectroscopy was used to determine the structure of these modified gA analogues and [Lys16]gramicidin A (gA-Lys) in sodium dodecyl-d25 sulphate micelles. Gated gA ion channels based on linking a receptor group to these gA analogues have been used recently as a component in a sensing device. The conformations of the gA backbones and amino acid side chains of lysinated gA and biotinylated gA in detergent micelles were found to be almost identical to that of native gA, i.e. that of an N-terminal to N-terminal (head to head) dimer formed by two right-handed, single-stranded beta 6.3 helices. The biotin tail of the gAXB and gAXXB and the lysine extremity of gA-Lys appeared to lie outside the micelle. Thus it appears that the covalent attachment of functional groups to the C terminus of gA does not disrupt the peptide's helical configuration. Further, single channel measurements of all three gA analogues showed that functioning ion channels were preserved within a membrane environment.
authors.affiliation: School of Chemistry, University of Melbourne, Parkville, VIC 3052, Australia. f.separovic@chemistry.unimelb.edu.au
authors.forename: F
authors.initials: F
authors.lastname: Separovic
authors.forename: S
authors.initials: S
authors.lastname: Barker
authors.forename: M
authors.initials: M
authors.lastname: Delahunty
authors.forename: R
authors.initials: R
authors.lastname: Smith
mesh_terms: D001710:Biotin / Q000737:chemistry; D006096:Gramicidin / Q000031:analogs & derivatives / Q000737:chemistry*; D007473:Ion Channels / Q000737:chemistry*; D008239:Lysine / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D011487:Protein Conformation; D011522:Protons; D012984:Software
chemical_list: D007473:Ion Channels; D011522:Protons; D006096:Gramicidin; D001710:Biotin; D008239:Lysine