Solubilization of human erythrocyte membranes by ASB detergents.
Understanding the membrane solubilization process and finding effective solubilizing agents are crucial challenges in biochemical research. Here we report results on the interaction of the novel linear alkylamido propyl dimethyl amino propanosulfonate detergents, ASB-14 and ASB-16, with human erythrocyte membranes. An estimation of the critical micelle concentration of these zwitterionic detergents (ASB-14 = 100 microM and ASB-16 = 10 microM) was obtained using electron paramagnetic resonance. The amount of proteins and cholesterol solubilized from erythrocytes by these detergents was then determined. The hemolytic activities of the ASB detergents were assayed and the detergent/lipid molar ratios for the onset of hemolysis (Re sat) and total lysis (Re sol) were calculated, allowing the determination of the membrane binding constants (Kb). ASB-14 presented lower membrane affinity (Kb = 7050 M(-1)) than ASB-16 (Kb = 15610 M(-1)). The amount of proteins and cholesterol solubilized by both ASB detergents was higher while Re sat values (0.22 and 0.08 detergent/lipid for ASB-14 and ASB-16, respectively) were smaller than those observed with the classic detergents CHAPS and Triton X-100. These results reveal that, besides their well-known use as membrane protein solubilizers to enhance the resolution of two dimensional electrophoresis/mass spectrometry, ASB-14 and ASB-16 are strong hemolytic agents. We propose that the physicochemical properties of ASB detergents determine their membrane disruption efficiency and can help to explain the improvement in the solubilization of membrane proteins, as reported in the literature.
Domingues
C C
CC
Departamento de Bioquímica, Instituto de Biologia, Universidade Estadual de Campinas, Campinas, SP, Brasil.
Malheiros
S V P
SV
Paula
E de
Ed
D017738:Alkanesulfonic Acids / Q000494:pharmacology*; D001622:Betaine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002784:Cholesterol / Q000378:metabolism*; D003902:Detergents / Q000494:pharmacology*; D004578:Electron Spin Resonance Spectroscopy; D015180:Electrophoresis, Gel, Two-Dimensional; D004910:Erythrocyte Membrane / Q000187:drug effects* / Q000378:metabolism; D006461:Hemolysis; D006801:Humans; D013058:Mass Spectrometry; D012995:Solubility
D017738:Alkanesulfonic Acids; D003902:Detergents; C506587:amidosulfobetaine-14; D001622:Betaine; D002784:Cholesterol