Abstract
A family of fluorescent probes, consisting of 2-p-toluidinylnapththalene-6-sulfonate (TNS) and neutral and cationic sulfonamido derivatives has been utilized to study the influence of electrostatic forces in protein-amphiphile interactions. 2-p-Toluidinylnaphthalene-6-[N-β-ethylammonium chloride] sulfonamide (III) binds to a lower number of discrete sites in bovine serum albumin and sperm whale apomyoglobin than does TNS, and is also bound less efficiently by β-lactoglobulin. The fluorescence characteristics of the bound probes indicate that their environments are hydrophobic, but thatpH and ionic strength influence the binding. The initial binding of III to discrete sites on both apomyoglobin and bovine serum albumin induces the cooperative binding of additional probe molecules. TNS, but not III, fluoresces in α-chymotrypsin solutions. An [N-β-ethyltrimethyl ammonium] sulfonamido derivative (IV), but not TNS, fluoresces in bovine trypsin solutions. Fluorescence-enhancing interactions were detected between TNS, III, and polyvinylpyrrolidone, but not between these probes and ribonuclease A, α-chymotrypsinogen, lysozyme, or γ-globulins. The wheat prolamin A-gliadin binds more TNS than III. The accessibility of all binding sites of gliadin is lower atpH 5.0 than atpH 3.1. It is suggested that, in general, charge effects are more likely to enhance the binding of anionic than cationic amphibiles by proteins.
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Dedicated to my teacher, Prof. Robert E. Feeney, on the occasion of his 70th Birthday. Presented in part at the Symposium on Chemical Modification and Structure-Function Relationships of Macromolecules, Division of Agricultural and Food Chemistry, 186th National Meeting of the American Chemical Society, 30 August 1983, Washington, D.C.
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Greene, F.C. Interactions of anionic and cationic fluorescent probes with proteins: The effect of charge. J Protein Chem 3, 167–180 (1984). https://doi.org/10.1007/BF01040498
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DOI: https://doi.org/10.1007/BF01040498