Abstract
Fluorescence quenching techniques have been used extensively in recent years to examine reaction rates and the compartmentalization of components in lipid micelles and membranes. Steady-state fluorescence methods are frequently employed in such studies but the interpretation of the resulting Stern-Volmer plots is often hampered by uncertainties regarding the mode of association of the quencher with the lipid structure and the nature of the quenching mechanism. This paper presents a method for simulating steady-state Stern-Volmer plots in two phase systems, and shows how the forms of such plots are influenced by the type of association of the quencher with the membrane or micelle (partition and/or binding) and by the type of quenching mechanism (dynamic and/or static). Comparisons of simulated plots with experimental data must take into account the possible combinations of quencher association(s) and quenching mechanism(s). The methods presented are applicable to synthetic and natural membranes and provide a basis for comparing the quenching of fluorescent molecules in biological membranes of differing composition.
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Selected References
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- Bieri V. G., Wallach D. F. Variations of lipid-protein interactions in erythrocyte ghosts as a function of temperature and pH in physiological and non-physiological ranges. A study using a paramagnetic quenching of protein fluorescence by nitroxide lipid analogues. Biochim Biophys Acta. 1975 Oct 17;406(3):415–423. doi: 10.1016/0005-2736(75)90020-6. [DOI] [PubMed] [Google Scholar]
- Eftink M. R., Ghiron C. A. Exposure of tryptophanyl residues and protein dynamics. Biochemistry. 1977 Dec 13;16(25):5546–5551. doi: 10.1021/bi00644a024. [DOI] [PubMed] [Google Scholar]
- Eftink M. R., Ghiron C. A. Exposure of tryptophanyl residues in proteins. Quantitative determination by fluorescence quenching studies. Biochemistry. 1976 Feb 10;15(3):672–680. doi: 10.1021/bi00648a035. [DOI] [PubMed] [Google Scholar]
- Haigh E. A., Thulborn K. R., Nichol L. W., Sawyer W. H. Uptake of n-(9-anthroyloxy) fatty acid fluorescent probes into lipid bilayers. Aust J Biol Sci. 1978 Oct;31(5):447–457. doi: 10.1071/bi9780447. [DOI] [PubMed] [Google Scholar]
- Klotz I. M., Hunston D. L. Properties of graphical representations of multiple classes of binding sites. Biochemistry. 1971 Aug 3;10(16):3065–3069. doi: 10.1021/bi00792a013. [DOI] [PubMed] [Google Scholar]
- Lakowicz J. R., Hogen D., Omann G. Diffusion and partitioning of a pesticide, lindane, into phosphatidylcholine bilayers. A new fluorescence quenching method to study chlorinated hydrocarbon-membrane interactions. Biochim Biophys Acta. 1977 Dec 15;471(3):401–411. doi: 10.1016/0005-2736(77)90045-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Omann G. M., Lakowicz J. R. Interactions of chlorinated hydrocarbon insecticides with membranes. Biochim Biophys Acta. 1982 Jan 4;684(1):83–95. doi: 10.1016/0005-2736(82)90052-9. [DOI] [PubMed] [Google Scholar]
- Shinitzky M., Dianoux A. C., Gitler C., Weber G. Microviscosity and order in the hydrocarbon region of micelles and membranes determined with fluorescent probes. I. Synthetic micelles. Biochemistry. 1971 May 25;10(11):2106–2113. doi: 10.1021/bi00787a023. [DOI] [PubMed] [Google Scholar]
- Wallach D. F., Verma S. P., Weidekamm E., Bieri V. Hydrophobic binding sites in bovine serum albumin and erythrocyte ghost proteins. Study by spin-labelling, paramagnetic fluorescence quenching and chemical modification. Biochim Biophys Acta. 1974 Jul 12;356(1):68–81. doi: 10.1016/0005-2736(74)90294-6. [DOI] [PubMed] [Google Scholar]