Fluorescence anisotropy measurements under oxygen quenching conditions as a method to quantify the depolarizing rotations of fluorophores. Application to diphenylhexatriene in isotropic solvents and in lipid bilayers

Lakowicz, Joseph R.; Prendergast, F. G.; Hogen, D.

doi:10.1021/bi00570a022

Cited by 110 publications

(65 citation statements)

References 34 publications

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“…The quenching of pyrene has been studied by Fischkoff and Vanderkooi in DMPC vesicles at 20~ [3]. The conclusion of this work is that oxygen can quench pyrene fluorescence in the membrane, and the derived diffusional coefficient for oxygen was 1.51 x 10 .5 cm2/s for DMPC at 37~ and 0.932 x 10 -5 cm2/s for DPPC at 37~ For TNS and DPH a similar conclusion was also reached by Lakowicz [1].…”

supporting

confidence: 76%

“…[1], have been measured in the same samples using emission wavelengths of 440 and 490 nm and excitation wavelengths of 340 or 410 nm (Fig. 4).…”

Section: Resultsmentioning

confidence: 99%

“…The localization and transport of molecular oxygen in the cell are of primary importance for the cell metabolism. Several groups have addressed the problem of determining the oxygen concentration and diffusion in the lipid bilayer [1][2][3][4][5][6][7][8]. There is a general consensus that oxygen solubility in the lipid phase is higher than in water [2].…”

mentioning

confidence: 99%

“…Among the methods used to measure oxygen solubility and diffusion is fluorescence oxygen quenching [1,3,9]. In a fluorescence quenching experiment, the results are generally analyzed using the Stern-Volmer quenching plot.…”

mentioning

confidence: 99%

“…Generally, the interpretation of the fluorescence experiment proceeds via an assumption about oxygen solubility, necessary for determining the abscissa of the Stern-Volmer plot, followed by calculation of the diffusion coefficient from its slope. Among the probes used for fluorescence quenching experiments in membranes, are pyrene [3], TNS, and DPH [1]. The quenching of pyrene has been studied by Fischkoff and Vanderkooi in DMPC vesicles at 20~ [3].…”

mentioning

confidence: 99%

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Packing of phospholipid vesicles studied by oxygen quenching of Laurdan fluorescence

Parasassi

Gratton

1992

J Fluoresc

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Steady-state fluorescence oxygen quenching experiments were performed on phospholipid vesicles where 2-dimethylamino-6-1auroylnaphthalene (Laurdan) was inserted. The quenching efficiency was found to be much higher in vesicles in the liquid-crystalline phase with respect to the gel phase, by a factor of about 50. Since the oxygen solubility in the two phospholipid phases can differ at most by a factor of 4 based on literature values, we concluded that oxygen diffusion must be responsible for the great difference in the quenching efficiency. A relatively high quenching efficiency was also found in vesicles composed of equimolar gel and liquid-crystalline phospholipids. Simulations were performed using the linear superposition of the properties of the pure phases to demonstrate that, in the case of vesicles composed of coexisting phases, the diffusional properties of oxygen in each phase are largely modified by the presence of the other. The addition of 10 mol% cholesterol to the gel phase rendered Laurdan fluorescence approximately as quenchable as in the equimolar mixture of the two phases. This result points out that molecules such as cholesterol, which introduce packing defects in the bilayer, favor oxygen diffusion. From the oxygen quenching experiments and using the properties of generalized polarization, the rate of Laurdan dipolar relaxation can be estimated.

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supporting

confidence: 76%

“…[1], have been measured in the same samples using emission wavelengths of 440 and 490 nm and excitation wavelengths of 340 or 410 nm (Fig. 4).…”

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Packing of phospholipid vesicles studied by oxygen quenching of Laurdan fluorescence

Parasassi

Gratton

1992

J Fluoresc

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Fluidity and Function of Hepatocyte Plasma Membranes

Schachter

1984

Hepatology

232

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LIPID-PROTEIN INTERACTIONS INMEMBRANES This review considers the basic biological problem of how the lipids and proteins of hepatocyte plasma membranes interact to mediate and regulate membrane functions. The following general hypothesis, which is applicable to all natural membranes, is a suitable framework for the discussion, inasmuch as it rests on a considerable body of experimental evidence. MEMBRANE MEMBRANE PROTEINS influence LIPIDS(and the specific -(and their functions properties) mediated by them)The influence of endogenous membrane enzymes, such as phospholipases (1-4), methyltransferases (5, 6), and fatty acid desaturases (7-lo), on membrane phospholipids is well-documented. Such reactions can alter the composition and properties of the bilayer lipid environment and thereby affect the functions of those membrane proteins ("integral" proteins) which are embedded in or traverse the lipid core of the membrane. A key property of the lipids, in this regard, is the "fluidity", or general motional freedom of the lipid molecules. A growing body of evidence indicates that the fluidity of the lipid environment influences the activity of such physiologically important membrane enzymes as the (Na+ + Ka+)-dependent adenosine triphosphatase (11-16) and hormoneresponsive adenylate cyclase (16-20). The fluidity also modulates transmembrane transport processes (21-24) and is a determinant of the passive permeability characteristic of a given bilayer (23, 25-28).The interactions of membrane proteins and lipids implicit in the general hypothesis above could provide regulatory loops for the control of important cell functions. For example, recent evidence indicates that calcium ion

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Fluorescence Quenching by Stimulated Emission

Lakowicz

Gryczyński

2002

Topics in Fluorescence Spectroscopy

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Fluorescence anisotropy measurements under oxygen quenching conditions as a method to quantify the depolarizing rotations of fluorophores. Application to diphenylhexatriene in isotropic solvents and in lipid bilayers

Cited by 110 publications

References 34 publications

Packing of phospholipid vesicles studied by oxygen quenching of Laurdan fluorescence

Packing of phospholipid vesicles studied by oxygen quenching of Laurdan fluorescence

Fluidity and Function of Hepatocyte Plasma Membranes

Fluorescence Quenching by Stimulated Emission

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