Immobilization and characterization of the transmembrane ion channel peptide gramicidin in a sol-gel matrix

Rocío Esquembre; José Antonio Poveda; Ricardo Mallavia; C. Reyes Mateo

doi:10.1117/12.721676

22 May 2007 Immobilization and characterization of the transmembrane ion channel peptide gramicidin in a sol-gel matrix

Rocío Esquembre, José Antonio Poveda, Ricardo Mallavia, C. Reyes Mateo

Proceedings Volume 6592, Bioengineered and Bioinspired Systems III; 659211 (2007) https://doi.org/10.1117/12.721676
Event: Microtechnologies for the New Millennium, 2007, Maspalomas, Gran Canaria, Spain

Abstract

Immobilization of ion channels requires of a methodology able to retain the physical properties of the lipid bilayer where their activity is performed. However, most of lipid membrane immobilization methods have been observed to alter the structural properties of the bilayers. Use of sol-gel routes seems to be an interesting alternative, although unstable liposomes were obtained when conventional sol-gel methodology was employed for immobilizing. Recently, we have suggested that use of alcohol-free sol-gel routes combined with negatively charged lipids could minimize effects exerted by host matrix on liposome structure, increasing its stability. Here we confirm this assumption by analysing the physical properties of a series of zwitterionic and anionic liposomes entrapped in a sol-gel matrix and we develop a methodology able to retain the physical properties of the lipid bilayer. This methodology has been successfully used to immobilize the transmembrane ion channel peptide gramicidin. Gramicidin was reconstituted in anionic liposomes and its immobilization was confirmed from changes observed in the photophysical properties of the tryptophan residues. Ion channel activity was determined using the fluorescent dye pyrene-1,3,6,8-tetrasulphonic acid (PTSA) and long term stability of the immobilized system was checked from steady-state fluorescence anisotropy measurements.

Citation Download Citation

Rocío Esquembre, José Antonio Poveda, Ricardo Mallavia, and C. Reyes Mateo "Immobilization and characterization of the transmembrane ion channel peptide gramicidin in a sol-gel matrix", Proc. SPIE 6592, Bioengineered and Bioinspired Systems III, 659211 (22 May 2007); https://doi.org/10.1117/12.721676

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