The most important physico-chemical characteristics describing detergent micelles are the CMC value and the aggregation number. These parameters depend critically on external conditions such as pH, ionic strength, buffer composition or temperature. However, the influence of these conditions on CMC or aggregation number is a priori not to predict, but the most important parameter. This holds especially for the aggregation number. Here, we present a simple, reliable and fast method for the analysis of detergent containing buffer systems. It enables the determination of CMC values and aggregation numbers of detergents as well as detergent mixtures in a 96well plate standard device relying on steady state fluorescence and fluorescence quenching. To prove the general applicability of our approach, we analyzed DDM as a prime example of a detergent and mixtures of DDM with six other detergents at different ratios to demonstrate the potential of our method.
KeywordsDetergentCMC DeterminationFluorescenceMixed MicellesAggregation Number
Jumpertz, T., Tschapek, B., Infed, N., Smits, S.H., Ernst, R. and Schmitt, L. (2011) High-Throughput Evaluation of the Critical Micelle Concentration of Detergents. Analytical Biochemistry, 408, 64-70. http://dx.doi.org/10.1016/j.ab.2010.09.011
Paradies, H.H. (1980) Shape and Size of a Nonionic Surfactant Micelle. Triton X-100 in Aqueous Solution. The Journal of Physical Chemistry, 84, 599-607. http://dx.doi.org/10.1021/j100443a008
Lund, S., Orlowski, S., de Foresta, B., Champeil, P., le Maire, M. and Moller, J.V. (1989) Detergent Structure and Associated Lipid as Determinants in the Stabilization of Solubilized Ca2+-ATPase from Sarcoplasmic Reticulum. The Journal of Biological Chemistry, 264, 4907-4915.
Infed, N., Hanekop, N., Driessen, A.J., Smits, S.H. and Schmitt, L. (2011) Influence of Detergents on the Activity of the ABC Transporter LmrA. Biochimica et Biophysica Acta, 1808, 2313-2321. http://dx.doi.org/10.1016/j.bbamem.2011.05.016
Screpanti, E., Padan, E., Rimon, A., Michel, H. and Hunte, C. (2006) Crucial Steps in the Structure Determination of the Na+/H+ Antiporter NhaA in Its Native Conformation. Journal of Molecular Biology, 362, 192-202. http://dx.doi.org/10.1016/j.jmb.2006.07.019
Lemieux, M.J., Song, J., Kim, M.J., Huang, Y., Villa, A., Auer, M., Li, X. D. and Wang, D.N. (2003) Three-Dimensional Crystallization of the Escherichia coli Glycerol-3-Phosphate Transporter: A Member of the Major Facilitator Superfamily. Protein Science, 12, 2748-2756. http://dx.doi.org/10.1110/ps.03276603
Michel, H. (1983) Crystallization of Membrane Proteins. Trends in Biochemical Sciences, 8, 56-59. http://dx.doi.org/10.1016/0968-0004(83)90390-0
Tummino, P.J. and Gafni, A. (1993) Determination of the Aggregation Number of Detergent Micelles Using Steady-State Fluorescence Quenching. Biophysical Journal, 64, 1580-1587. http://dx.doi.org/10.1016/S0006-3495(93)81528-5
Turro, N.J. and Yekta, A. (1978) Luminescent Probes for Detergent Solutions. A simple Procedure for Determination of Mean Aggregation Number of Micelles. Journal of the American Chemical Society, 100, 5951-5952. http://dx.doi.org/10.1021/ja00486a062
Infelta, P.P. (1978) Fluorescence Quenching in Micellar Solutions and Its Application to the Determination of Aggregation Numbers. Chemical Physics Letters, 61, 88-91. http://dx.doi.org/10.1016/0009-2614(79)85092-7
Prive, G.G. (2007) Detergents for the Stabilization and Crystallization of Membrane Proteins. Methods, 41, 388-397. http://dx.doi.org/10.1016/j.ymeth.2007.01.007
Pastor, O., Junquera, E. and Aicart, E. (1998) Hydration and Micellization Processes of n-Octyl-β-D-Glucopyranoside in Aqueous Solution. A Thermodynamic and Fluorimetric Study in the Absence and Presence of Salts. Langmuir, 14, 2950-5957. http://dx.doi.org/10.1021/la9708445
Atik, S.S. and Singer, L.A. (1978) Nitroxyl Radical Quenching of the Pyrene Fluorescence in Micellar Environments. Development of a Kinetic Model for Steady-State and Transient Experiments. Chemical Physics Letters, 59, 519-524. http://dx.doi.org/10.1016/0009-2614(78)85032-5
Hansson, P., Jönsson, B., Ström, C. and Sdermann, O. (2000) Determination of Micellar Aggregation Numbers in Dilute Surfactant Systems with the Fluorescence Quenching Method. The Journal of Physical Chemistry B, 104, 3496-3506. http://dx.doi.org/10.1021/jp992444r
Clint, J.H. (1974) Micellization of Mixed Nonionic Surface Active Agents. Journal of the Chemical Society, Faraday Transactions, 71, 1327-1334. http://dx.doi.org/10.1039/f19757101327
Sarmoria, C., Puvvada, S. and Blankschtein, D. (1992) Prediction of Critical Micelle Concentrations of Nonideal Binary Surfactant Mixtures. Langmuir, 8, 2690-2697. http://dx.doi.org/10.1021/la00047a019
Lipfert, J., Columbus, L., Chu, V.B., Lesley, S.A. and Doniach, S. (2007) Size and Shape of Detergent Micelles Determined by Small-Angle X-Ray Scattering. The Journal of Physical Chemistry B, 111, 12427-12438. http://dx.doi.org/10.1021/jp073016l
Bezzobotnov, V.Y., Borbély, S., Cser, L., Faragó, B., Gladkih, I.A., Ostanevich, Y.M. and Vass, S. (1988) Temperature and Concentration Dependence of Properties of Sodium Dodecyl Sulfate Micelles Determined from Small Angle Neutron Scattering Experiments. The Journal of Physical Chemistry, 92, 5738-5743. http://dx.doi.org/10.1021/j100331a038
Gangabadage, C.S., Najda, A., Bogdan, D., Wijmenga, S.S. and Tessari, M. (2008) Dependence of the Size of a Protein-SDS Complex on Detergent and Na+ Concentrations. The Journal of Physical Chemistry B, 112, 4242-4245. http://dx.doi.org/10.1021/jp710045e
Bucci, S., Fagotti, C., Degiorgio, V. and Piazza, R. (1991) Small-Angle Neutron-Scattering Study of Ionic-Nonionic Mixed Micelles. Langmuir, 7, 824-826. http://dx.doi.org/10.1021/la00053a002
Doughty, D.A. (1979) Isopiestic Compositions of Aqueous Ionic Surfactant Systems as a Measure of Preferential Interactions. Application to the Determination of Micelle Aggregation Numbers by Equilibrium Ultracentrifugation. The Journal of Physical Chemistry, 83, 2621-2628. http://dx.doi.org/10.1021/j100483a014
Lianos, P. and Zana, R. (1980) Use of Pyrene Excimer Formation to Study the Effect of Sodium Chloride on the Structure of Sodium Dodecyl Sulfate Micelles. The Journal of Physical Chemistry, 84, 3339-3341. http://dx.doi.org/10.1021/j100462a003
Emerson, M.F. and Holtzer, A. (1967) On the Ionic Strength Dependence of Micelle Number II. The Journal of Physical Chemistry, 71, 1898-1907. http://dx.doi.org/10.1021/j100865a057
Bales, B.L., Messina, L., Vidal, A. and Peric, M. (1998) Precision Relative Aggregation Number Determinations of SDS Micelles Using a Spin Probe. A Model of Micelle Surface Hydration. The Journal of Physical Chemistry B, 102, 10347-10358. http://dx.doi.org/10.1021/jp983364a
Motomura, K., Yamanaka, M. and Aranato, M. (1984) Thermodynamic Consideration of the Mixed Micelle of Surfactants. Colloid & Polymer Science, 262, 948-955. http://dx.doi.org/10.1007/BF01490027
Zhang, R., Zhang, L. and Somasundaran, P. (2003) Study of Mixtures of n-Dodecyl-β-D-Maltoside with Anionic, Cationic, and Nonionic Surfactant in Aqueous Solutions Using Surface Tension and Fluorescence Techniques. Journal of Colloid and Interface Science, 278, 453-460. http://dx.doi.org/10.1016/j.jcis.2004.06.045
Columbus, L., Lipfert, J., Jambunathan, K., Fox, D.A., Sim, A.Y., Doniach, S. and Lesley, S.A. (2009) Mixing and Matching Detergents for Membrane Protein NMR Structure Determination. Journal of the American Chemical Society, 131, 7320-7326. http://dx.doi.org/10.1021/ja808776j
Turro, N.J., Graetzle, M. and Braun, A.M. (1980) Photophysical and Photochemical Processes in Micellar Systems. Angewandte Chemie International Edition in English, 19, 675-696. http://dx.doi.org/10.1002/anie.198006751
Herrmann, K.W. (1962) Non-Ionic-Cationic Micellar Properties of Dimethyldodecylamine Oxide. The Journal of Physical Chemistry, 66, 295-300. http://dx.doi.org/10.1021/j100808a025
Herrmann, K.W. (1966) Micellar Properties of Some Zwitterionic Surfactants. Journal of Colloid and Interface Science, 22, 352-359. http://dx.doi.org/10.1016/0021-9797(66)90015-4