Raft composition at physiological temperature and pH in the absence of detergents.

摘要: Biological rafts were identified and isolated at 37°C neutral pH. The strategy for isolating utilized membrane tension to generate large domains. For lipid compositions that led only microscropically unresolvable in bilayers, the appearance of large, observable rafts. converted back small ones when was relieved. Thus, reversibly controls raft enlargement. cells, application resulted several types domains; one class domains as Tension generated ways, all yielded fractions had essentially same composition, validating principle a means merge into It demonstrated sphingomyelin-rich vesicles do not rise during centrifugation sucrose gradients because they resist lysis, necessitating that, contrary current experimental practice, material be placed toward top gradient fractionation. Isolated enriched GPI-linked protein, alkaline phosphatase, poor Na+-K+ ATPase. Sphingomyelin gangliosides concentrated rafts, expected composition. Cholesterol, however, distributed equally between nonraft fractions, conventional view.