NDLI: Photoactivable sphingosine as a tool to study membrane microenvironments in cultured cells.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Aureli, Massimo Prioni, Simona Mauri, Laura Loberto, Nicoletta Casellato, Riccardo Ciampa, Maria Grazia Chigorno, Vanna Prinetti, Alessandro Sonnino, Sandro
Description	Country affiliation: Italy Author Affiliation: Aureli M ( Department of Medical Chemistry Biochemistry and Biotechnology Center of Excellence on Neurodegenerative Diseases, University of Milano, Segrate, Italy.)
Abstract	Human fibroblasts from normal subjects and Niemann-Pick A (NPA) disease patients were fed with two labeled metabolic precursors of sphingomyelin (SM), [(3)H]choline and photoactivable sphingosine, that entered into the biosynthetic pathway allowing the synthesis of radioactive phosphatidylcholine and SM, and of radioactive and photoactivable SM ([(3)H]SM-N(3)). Detergent resistant membrane (DRM) fractions prepared from normal and NPA fibroblasts resulted as highly enriched in [(3)H]SM-N(3). However, lipid and protein analysis showed strong differences between the two cell types. After cross-linking, different patterns of SM-protein complexes were found, mainly associated with the detergent soluble fraction of the gradient containing most cell proteins. After cell surface biotinylation, DRMs were immunoprecipitated using streptavidin. In conditions that maintain the integrity of domain, SM-protein complexes were detectable only in normal fibroblasts, whereas disrupting the membrane organization, these complexes were not recovered in the immunoprecipitate, suggesting that they involve proteins belonging to the inner membrane layer. These data suggest that differences in lipid and protein compositions of these cell lines determine specific lipid-protein interactions and different clustering within plasma membrane. In addition, our experiments show that photoactivable sphingolipids metabolically synthesized in cells can be used to study sphingolipid protein environments and sphingolipid-protein interactions.
File Format	HTM / HTML
ISSN	00222275
e-ISSN	15397262
DOI	10.1194/jlr.M001974
Journal	The Journal of Lipid Research
Issue Number	4
Volume Number	51
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	2010-04-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Biochemistry Cell Membrane Metabolism Membrane Microdomains Sphingomyelins Biosynthesis Sphingosine Analogs & Derivatives Biotinylation Cell Proliferation Cell Survival Cells, Cultured Cross-linking Reagents Immunoprecipitation Lipids Membrane Proteins Niemann-pick Disease, Type A Phosphatidylcholines Photochemical Processes Skin Cytology Adverse Effects Chemical Synthesis Streptavidin Time Factors Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Cell Biology Biochemistry Endocrinology

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Photoactivable sphingosine as a tool to study membrane microenvironments in cultured cells[S]

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Photoactivable sphingosine as a tool to study membrane microenvironments in cultured cells.

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Photoactivable sphingosine as a tool to study membrane microenvironments in cultured cells[S]

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Photoactivable sphingosine as a tool to study membrane microenvironments in cultured cells.