Cooperative binding of hexadecyltrimethylammonium chloride and sodium dodecyl sulfate to cytochrome c and the resultant change in protein conformation.

The binding of hexadecyltrimethylammonium chloride (HTAC) and sodium dodecyl sulfate (SDS) to cytochrome c was determined by potentiometric titration and the corresponding changes in protein conformation by circular dichroism (CD). The binding isotherms were biphasic; about 20 surfactant cations or anions were bound to cytochrome c in the first phase. Another 30 or so HTA+ ions were bound in the second phase, which was below the critical micelle concentration of the surfactant, but the binding of dodecyl sulfate ions in the second phase increased sharply near the critical micelle concentration. The binding of both surfactants was highly cooperative and was endothermic; the data in the first phase fitted the Hill plot. The corresponding change in the secondary structure of cytochrome c was small; the CD spectra in the ultraviolet region showed a moderate increase in the helicity in HTAC solution and some changes in conformation in SDS solution. However, the CD spectra for the Soret band indicated a marked change in the local conformation around the heme.