Process of Destruction of Large Unilamellar Vesicles by a Zwitterionic Detergent, CHAPS: Partition Behavior between Membrane and Water Phases

Abstract

The process of vesicle destruction by zwitterionic detergent, 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), was examined to clarify the vesicle-micelle transition mechanism. The physicochemical properties including turbidity, apparent particle size, Cl⁻ permeability, electron spin resonance (ESR) spectroscopic parameters, and freeze-fracture electron microscopy were investigated. The concentration of CHAPS was analyzed using HPLC to determine the partition coefficient during the solubilization process. The data obtained revealed that maximum turbidity and apparent particle size were found at the effective ratio (R_e) of 0.21 and 0.49, respectively. With a further increase in CHAPS concentration, turbidity and particle size abruptly decreased, suggesting the formation of mixed micelles. The partition coefficient changed throughout the solubilization process. In the presence of low concentrations of CHAPS, CHAPS partitioned into vesicles without destruction of membrane bilayers. When the R_e<0.04, the partition coefficient was independent of the detergent concentration with value of 24 M⁻¹. At R_e greater than 0.05, the membrane barrier abruptly decreased. At 0.04≤R_e<0.21, the gradual increase in the partition coefficient accounted for the occurrence of larger vesicles. In range of 0.21≤R_e<0.52, the abrupt increase in the partition behavior was possibly attributed to the structural change of mixed vesicles to mixed micelles. Furthermore, the ESR results showed that the incorporation of CHAPS into vesicles led to an increase in membrane fluidity near the polar head, and a decrease near the end of the acyl chain. ESR spectra of 5-doxylstearic acid in CHAPS-containing micelles were anisotropic, indicating that the steroidal structure of CHAPS was responsible for the micelles possessing an orderly arrangement of hydrocarbon chains.