Chromatophores isolated from cells of Rhodobacter sphaeroides exposed to hypertonic solutions were enriched in cardiolipin (CL). As CL raises by increasing the incubation time of R. sphaeroides in hypertonic solutions, it was possible to isolate chromatophores containing different CL amounts by starting from cells incubated in hypertonic solutions for different times. The functionality and stability of the photosynthetic proteins in chromatophore membranes having different CL levels were investigated. Reaction Center (RC) stabilization with respect to thermal denaturation and photoxidative damage was observed by flash photolysis and fluorescence emission experiments in CL enriched chromatophores. In order to gain detailed information about the structures of endogenous CLs, this lipid family was isolated and purified by preparative TLC, and characterized by high resolution mass spectrometry. We conclude that osmotic shock can be used as a tool to modulate CL levels in isolated chromatophores and to change the composition of the RC lipid annulus, avoiding membrane artefacts introduced by the use of detergents.
Cardiolipin increases in chromatophores isolated from Rhodobacter sphareroides after osmotic stress: structural and functional roles
DE LEO V;CATUCCI, Lucia;AGOSTIANO, Angela;CORCELLI, Angela
2009-01-01
Abstract
Chromatophores isolated from cells of Rhodobacter sphaeroides exposed to hypertonic solutions were enriched in cardiolipin (CL). As CL raises by increasing the incubation time of R. sphaeroides in hypertonic solutions, it was possible to isolate chromatophores containing different CL amounts by starting from cells incubated in hypertonic solutions for different times. The functionality and stability of the photosynthetic proteins in chromatophore membranes having different CL levels were investigated. Reaction Center (RC) stabilization with respect to thermal denaturation and photoxidative damage was observed by flash photolysis and fluorescence emission experiments in CL enriched chromatophores. In order to gain detailed information about the structures of endogenous CLs, this lipid family was isolated and purified by preparative TLC, and characterized by high resolution mass spectrometry. We conclude that osmotic shock can be used as a tool to modulate CL levels in isolated chromatophores and to change the composition of the RC lipid annulus, avoiding membrane artefacts introduced by the use of detergents.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.