Biophysical and functional characterization of an ion channel peptide confined in a sol-gel matrix.
Immobilization of zwitterionic lipid membranes in sol-gel matrices induces irreversible alterations of the bilayer fluidity, which can limit the use of these systems for practical applications. Recently, we have reported that electrostatic interactions between phospholipids polar heads and the negative-charged silica surface of the porous matrix should be the cause of such behavior. In the present work, we analyze the effect of these interactions on the biophysical and functional properties of the ion-channel peptide gramicidin, entrapped in a sol-gel matrix, to get more insight on the ability of these inorganic materials to immobilize ion channels and other membrane-bound proteins. Gramicidin was reconstituted in anionic and zwitterionic liposomes and the effects of sol-gel immobilization on the biophysical properties of gramicidin were determined from changes in the photophysical properties of its tryptophan residues. In addition, the physical state of the immobilized lipid membrane containing gramicidin was analyzed by measuring the spectral shift of the fluorescent probe Laurdan. Finally, the ion-channel activity of the peptide was monitored upon sol-gel immobilization through a fluorescence quenching assay using the fluorescent dye pyrene-1,3,6,8-tetrasulfonic acid (PTSA). Results show that the channel properties of the immobilized gramicidin are preserved in both zwitterionic and anionic liposomes, even though the zwitterionic polar heads interact with the porous surface of the host matrix.
Esquembre
Rocío
R
Instituto de Biología Molecular y Celular, Universidad Miguel Hernández de Elche, 03202 Elche, Alicante, Spain.
Poveda
José Antonio
JA
Mateo
C Reyes
CR
D055592:Biophysical Phenomena; D002412:Cations / Q000737:chemistry; D002586:Cesium / Q000737:chemistry; D005456:Fluorescent Dyes; D005782:Gels / Q000737:chemistry*; D006096:Gramicidin / Q000737:chemistry*; D007473:Ion Channels / Q000737:chemistry*; D008081:Liposomes; D008567:Membranes, Artificial; D017646:Organosilicon Compounds / Q000737:chemistry*; D010713:Phosphatidylcholines / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
D002412:Cations; D005456:Fluorescent Dyes; D005782:Gels; D007473:Ion Channels; D008081:Liposomes; D008567:Membranes, Artificial; D017646:Organosilicon Compounds; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D010743:Phospholipids; C500739:tetramethyl orthosilicate; D006096:Gramicidin; D002586:Cesium