gramicidin-a and phthalocyanine

Interaction of potent photodynamic agents, sulfonated aluminum phthalocyanines (AlPcSn where n is a number of sulfonic groups), with biological membranes was studied here using model systems: sensitized photoinactivation of gramicidin channels in planar lipid bilayers and adsorption on lipid monolayers. Fluoride anions known to form complexes with aluminum were found to inhibit both the adsorption of aluminum phthalocyanines on lipid monolayers, as measured with a Langmuir trough by surface pressure and surface potential changes, and photodynamic efficacy of the dyes, as studied by gramicidin channel photoinactivation. The similar effects were caused by the alkalinization of the medium. Fluoride anions appeared to be much more effective in the case of AlPcS4 as compared to AlPcS3. The suppression of the photodynamic potency of aluminum phthalocyanines was attributed to desorption of the dyes from lipid bilayers induced by fluoride or hydroxyl ions. With AlPcS4 an enhancement of the dye aggregation leading to a decrease in the sensitizing activity was probably involved in the fluoride effect as revealed by absorption and fluorescence spectral measurements. Capillary electrophoresis was employed to understand the mechanism of the dye desorption. The results of these experiments indicated that the reduction in the membrane affinity was associated with an increase in the negative charge of the dye molecules due to the binding of fluoride or hydroxyl ions.

Topics: Fluorides; Gramicidin; Hydrogen-Ion Concentration; In Vitro Techniques; Indoles; Isoindoles; Lipid Bilayers; Photochemistry; Photochemotherapy; Photosensitizing Agents

Photodynamic inactivation of gramicidin channels in bilayer lipid membranes induced by single flashes of the visible light in the presence of phthalocyanine has been studied. The kinetic curves of the flash-induced decrease in the gramicidin-mediated electric current are used for determination of the rate constants of formation and termination of gramicidin channels in terms of the channel dimer model. It is revealed that the kinetics of the sensitized photoinactivation of gramicidin in the membrane is altered by agents which modify the dipole potential drop at the membrane-water interface. Addition of phloretin, which is known to decrease the dipole potential drop, slows down the kinetics, whereas the addition of RH421 or 6-ketocholestanol, which increase the dipole potential drop, accelerates the kinetics. It is shown that the photoinactivation kinetics is also slowed down upon the addition of the thyroid hormone L-thyronine, which reduces the dipole potential drop similar to phloretin, as it was found earlier (M. V. Tsybulskaya, Yu. N. Antonenko, A. E. Tropsha, and L. S. Yaguzhinsky, Biofizika 29:801-805 (1984) (in Russian)). It is demonstrated that the changes in the dissociation rate constant of gramicidin dimers under the action of different dipole modifiers correlate with the changes in the dipole potential drop. It is concluded that the process of the gramicidin channel termination corresponding to the dimer dissociation is sensitive to the dipole potential drop. This conclusion is supported by the data on the effect of dipole modifiers on the lifetime of single gramicidin channels.

Topics: Gramicidin; Indoles; Isoindoles; Ketocholesterols; Kinetics; Lipid Bilayers; Phloretin; Phosphatidylcholines; Pyridinium Compounds; Radiation-Sensitizing Agents; Styrenes; Thyronines

Photosensitized inactivation of ionic channels formed by gramicidin in the planar bilayer lipid membrane (BLM) has been studied upon exposure of the BLM to single flashes of visible light in the presence of tetrasulphonated aluminium phthalocyanine. The gramicidin photoinactivation is inhibited by the addition of unsaturated phospholipids to the membrane-forming solution as well as by the addition of azide to the bathing solution, consistent with involvement of singlet oxygen. The characteristic time of the photoinactivation (tau) does not change markedly under these conditions. Moreover, tau remains nearly constant upon alteration of the flash energy and the photosensitizer concentration. The value of tau appears to be sensitive to the gramicidin concentration and to the factors affecting the open time of the gramicidin channels, namely the temperature and the solvent used in the membrane-forming solution. The photoinactivation is not observed with covalent gramicidin dimers. The equations derived from the model of Bamberg and Laeuger (J. Membrane Biol. (1973) 11, 177-194), describing the relaxation of the gramicidin-induced conductance after a sudden distortion of the dimer-monomer equilibrium, are shown to explain consistently the time course of the photoinactivation provided that the damage of the gramicidin molecules leads to deviation from the equilibrium.

Topics: Gramicidin; Indoles; Ion Channels; Isoindoles; Lipid Bilayers; Membranes, Artificial; Photolysis

The effect of phthalocyanines, the potent photodynamic sensitizers, on the electric properties of the bilayer lipid membrane (BLM) is studied. It is shown, that tetrasulfonated, as well as trisulfonated, aluminium phthalocyanine do not alter the conductance of BLM, but elicit certain changes in the boundary potential difference, which points in favor of dye adsorption on BLM. Under the conditions of intense visible light irradiation, the phthalocyanines cause an increase in the conductance, resulting in the irreversible breakdown of BLM, formed from soy bean phosphatidylcholine, but fail to change the conductance of BLM, formed from diphytanoilphosphatidylcholine. The phthalocyanine-sensitized inactivation of gramicidin channels incorporated into BLM is observed under the conditions of weak visible light irradiation using an He-Ne laser. The photodynamic blockage of model ionic channels is considerably suppressed after oxygen depletion. The phenomenon consists of a marked reduction of a number of open channels, probably due to photomodification of tryptophan residues, essential for gramicidin functioning. The mechanism of the channel inactivation, involving the photosensitized reaction of the II type, and the relevance to the interaction of sensitizers with biomembranes, is discussed.

Topics: Gramicidin; Indoles; Ion Channels; Isoindoles; Light; Lipid Bilayers; Oxidation-Reduction; Photosensitizing Agents; Reactive Oxygen Species