title: Fluorescence study of the macrolide pentaene antibiotic filipin in aqueous solution and in a model system of membranes.
abstract: The polyene antibiotic filipin (a pentaene) has been studied using photophysical techniques. The polyene self-aggregates in water with a critical micellar concentration of 2 microM. Two approaches were used to evaluate the aggregate dimensions: (a) a lower limit of 10 nm for the aggregate radius was obtained from energy transfer experiments; (b) a formula for rationalizing the turbidity spectrum was derived, and from its application a spherical shape of radius about 50 nm was deduced. The low value for the fluorescence anisotropy of the aggregate (r = 0.02) is compatible with a very loose structure, i.e. the chromophore has very efficient depolarization dynamics that is not controlled by the aggregate size. The Stern-Volmer plot of aggregated filipin fluorescence quenching by iodide is non-linear, presenting a downward curvature. A model was used for the interpretation of these data, along with a study of the quenching in transient state; it was concluded that all the components of the decay are affected by the quencher, i.e. the aggregate has a very open structure with respect to the iodide ion. The partition constants of the polyene, Kp, between a model system of membranes (small unilamellar vesicles of dipalmitoylglycerophosphocholine) and the aqueous phase were determined from anisotropy measurements; the values obtained were Kp (gel phase) = (3.4 +/- 0.8) x 10(3) and Kp (liquid crystal phase) = (7.7 +/- 2.2) x 10(2). The observation that the polyene incorporation is efficient is at variance with the belief that the presence of sterols are essential for the interaction of polyene antibiotics with membranes [for review see Bolard, J. (1986) Biochim. Biophys. Acta 864, 257-304].
authors.affiliation: Centro de Química Fisica Molecular, Lisboa, Portugal.
authors.forename: M A
authors.initials: MA
authors.lastname: Castanho
authors.forename: M J
authors.initials: MJ
authors.lastname: Prieto
mesh_terms: D004735:Energy Transfer; D005372:Filipin / Q000737:chemistry*; D005454:Fluorescence Polarization; D007700:Kinetics; D008027:Light; D008081:Liposomes; D008433:Mathematics; D008962:Models, Theoretical; D008968:Molecular Conformation; D015394:Molecular Structure; D011789:Quantum Theory; D012542:Scattering, Radiation; D012996:Solutions; D013050:Spectrometry, Fluorescence / Q000379:methods; D014867:Water
chemical_list: D008081:Liposomes; D012996:Solutions; D014867:Water; D005372:Filipin