Laurdan and Prodan as Polarity-Sensitive Fluorescent Membrane Probes

The steady-state and dynamic fluorescence spectral properties of 2-dimethylamino-6-lauroylnaphthalene (LAURDAN) and several other naphthalene derivatives are summarized to illustrate their sensitivity to the polarity of the environment. Results obtained both in solvents of different polarity and in phospholipid vesicles in two phase states are presented. The emission red shift observed in polar solvents and in the phospholipid liquid–crystalline phase is explained on the basis of dipolar relaxation of solvent molecules surrounding the fluorescent naphthalene moiety of these probes. In phospholipid environments, experimental evidence is shown that excludes the intramolecular relative reorientation of the dimethylamino and carbonyl groups in the naphthalene and the reorientation of the entire fluorescent moiety. The solvent dipolar relaxation observed for LAURDAN and PRODAN in phospholipid bilayers has been attributed to a small number of water molecules present at the membrane interface. A comparison between LAURDAN emission in phospholipid vesicles prepared in D2O and in H2O is also presented. The definition and the derivation of the generalized polarization function are also discussed.

[1]  M. Paternostre,et al.  Laurdan solvatochromism: solvent dielectric relaxation and intramolecular excited-state reaction. , 1997, Biophysical journal.

[2]  E. Gratton,et al.  Evidence for an increase in water concentration in bilayers after oxidative damage of phospholipids induced by ionizing radiation. , 1994, International journal of radiation biology.

[3]  A. Santroni,et al.  Effect of aromatic nitroxides on hemolysis of human erythrocytes entrapped with isolated hemoglobin chains. , 1997, Free radical biology & medicine.

[4]  F. Young.  Biochemistry , 1955, The Indian Medical Gazette.

[5]  T. Parasassi,et al.  In K562 and HL60 cells membrane ageing during cell growth is associated with changes in cholesterol concentration , 1997, Mechanisms of Ageing and Development.

[6]  Enrico Gratton,et al.  MODULATION AND DYNAMICS OF PHASE PROPERTIES IN PHOSPHOLIPID MIXTURES DETECTED BY LAURDAN FLUORESCENCE * , 1993, Photochemistry and photobiology.

[7]  E Gratton,et al.  Fluorescence generalized polarization of cell membranes: a two-photon scanning microscopy approach. , 1996, Biophysical journal.

[8]  J. Lakowicz,et al.  Quenching of fluorescence by oxygen. A probe for structural fluctuations in macromolecules. , 1973, Biochemistry.

[9]  F. Barrantes,et al.  Nicotinic acetylcholine receptor channels are influenced by the physical state of their membrane environment. , 1996, Biophysical journal.

[10]  F. Barrantes,et al.  Physical state of bulk and protein-associated lipid in nicotinic acetylcholine receptor-rich membrane studied by laurdan generalized polarization and fluorescence energy transfer. , 1996, Biophysical journal.

[11]  E Gratton,et al.  Quantitation of lipid phases in phospholipid vesicles by the generalized polarization of Laurdan fluorescence. , 1991, Biophysical journal.

[12]  G. Weber,et al.  Synthesis and spectral properties of a hydrophobic fluorescent probe: 6-propionyl-2-(dimethylamino)naphthalene. , 1979, Biochemistry.

[13]  M. Ruonala,et al.  Evidence for superlattice arrangements in fluid phosphatidylcholine/phosphatidylethanolamine bilayers. , 1997, Biophysical journal.

[14]  L. Bagatolli,et al.  Laurdan properties in glycosphingolipid-phospholipid mixtures: a comparative fluorescence and calorimetric study. , 1997, Biochimica et biophysica acta.

[15]  J. D. Bell,et al.  Interactions of thionin from Pyrularia pubera with dipalmitoylphosphatidylglycerol large unilamellar vesicles. , 1997, Biochemistry.

[16]  E. Gratton,et al.  Absence of lipid gel-phase domains in seven mammalian cell lines and in four primary cell types. , 1993, Biochimica et biophysica acta.

[17]  G. Weber.  Polarization of the fluorescence of macromolecules. I. Theory and experimental method. , 1952, The Biochemical journal.

[18]  J. D. Bell,et al.  Enhancement of Agkistrodon piscivorus piscivorus venom phospholipase A2 activity toward phosphatidylcholine vesicles by lysolecithin and palmitic acid: studies with fluorescent probes of membrane structure. , 1995, Biochemistry.

[19]  P. Chong,et al.  Interactions of Laurdan with phosphatidylcholine liposomes: a high pressure FTIR study. , 1993, Biochimica et biophysica acta.

[20]  C. Sotomayor,et al.  Effect of polyunsaturated fatty acid deficiency on dipole relaxation in the membrane interface of rat liver microsomes. , 1997, Biochimica et biophysica acta.

[21]  T. Galeazzi,et al.  Effect of aluminium ions on liposomal membranes as detected by Laurdan fluorescence. , 1997, Free radical research.

[22]  G. Weber,et al.  FLUOROPHORES IN POLAR MEDIA: SPECTRAL EFFECTS OF THE LANGEVIN DISTRIBUTION OF ELECTROSTATIC INTERACTIONS , 1981 .

[23]  F. G. Prendergast,et al.  Singlet adiabatic states of solvated PRODAN: a semiempirical molecular orbital study , 1989 .

[24]  E. Gratton,et al.  Two-photon fluorescence microscopy of laurdan generalized polarization domains in model and natural membranes. , 1997, Biophysical journal.

[25]  W. Pryor,et al.  Effect of lipid ozonation products on liposomal membranes detected by Laurdan fluorescence. , 1995, Free radical biology & medicine.