Interaction of cationic detergents, cetyl- and dodecyl-trimethylammonium bromides, with lysozyme.

Abstract

Binding of lysozyme with cetyltrimethylammonium bromide (CTAB) and dodecyl-trimethylammonium bromide (DTAB) at various detergent concentrations and pH was studied at 25 degrees C by equilibrium dialysis technique. In the case of CTAB, binding isotherms at pH 5.0, 7.0, and 9.0 show cooperative binding at all the concentrations of the detergent and the binding ratios increase with pH. Cooperative binding is also shown by DTAB at all the concentrations and pH, but the binding ratios are lower compared to CTAB. The Gibb's free energy change calculated on the basis of Wyman's binding potential concept increases with pH, indicating increased binding strength of CTAB at higher pH. The UV difference spectra of CTAB and DTAB with lysozyme and its model compounds such as L-Trp, L-Tyr X HCI and L-Phe show two peaks at 297 nm and 250 nm at pH 9.0 indicating the possible involvement of tryptophans as the binding sites along with the carboxylate anion or the phenolic group of a tyrosine on lysozyme. The effect of higher ionic strength on the binding of CTAB with lysozyme at pH 9.0 is evidenced by lower binding ratios and decreased intensities of the UV difference bands, thus indicating the involvement of electrostatic interactions. However, the hydrophobic interactions between the detergents and the aromatic amino acid residues in lysozyme contribute more to the binding strength. The binding of these cationic detergents by lysozyme induces conformational changes in the enzyme. They are followed by the circular dichroism (CD) technique which shows a decrease in the aromatic bands in the 320-250 nm region.(ABSTRACT TRUNCATED AT 250 WORDS)