Sphingolipid-enriched membrane domains from rat cerebellar granule cells differentiated in culture. A compositional study

Alessandro Prinetti, Vanna Chigorno, Guido Tettamanti, Sandro Sonnino

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Abstract

Sphingolipid-enriched membrane domains, characterized by a particular protein and lipid composition, have been detected in a variety of cells. However, limited data are available concerning these domains in neuronal cells. We analyzed the lipid and protein composition of a sphingolipid- enriched membrane fraction prepared from primary rat cerebellar granule cells differentiated in culture. Although the protein content of this fraction was only 1.4% of total cellular protein, 60% of the gangliosides, 67% of the sphingomyelin, 50% of the ceramide, and 40% of the cholesterol were located in this fraction. The protein pattern of the sphingolipid-enriched domain fraction was dramatically different from that associated with the cell homogenate. This fraction contained 25% of the tyrosine-phosphorylated proteins and was enriched in two proteins with apparent molecular masses of 135 and 15 kDa. 12% of cellular glycerophospholipids were located in the fraction, with phosphatidylcholine having the highest enrichment. The molar ratio between proteins, glycerophospholipids, cholesterol, sphingomyelin, ceramide and gangliosides in cerebellar granule cells was 1.6:41.6:6.1:1.3:0.3:1 in the cell homogenate and 0.04:8.3:4.0:1.4:0.2:1 in the sphingolipid-enriched membrane fraction. These data indicate that selected proteins segregate with sphingolipids in specialized domains in the membrane of cultured neurons.

Original languageEnglish
Pages (from-to)11658-11665
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number16
DOIs
Publication statusPublished - Apr 21 2000

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Sphingolipids
Rats
Membranes
Proteins
Glycerophospholipids
Sphingomyelins
Gangliosides
Ceramides
Cholesterol
Lipids
Molecular mass
Chemical analysis
Phosphatidylcholines
Neurons
Tyrosine

ASJC Scopus subject areas

  • Biochemistry

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Sphingolipid-enriched membrane domains from rat cerebellar granule cells differentiated in culture. A compositional study. / Prinetti, Alessandro; Chigorno, Vanna; Tettamanti, Guido; Sonnino, Sandro.

In: Journal of Biological Chemistry, Vol. 275, No. 16, 21.04.2000, p. 11658-11665.

Research output: Contribution to journalArticle

Prinetti, Alessandro ; Chigorno, Vanna ; Tettamanti, Guido ; Sonnino, Sandro. / Sphingolipid-enriched membrane domains from rat cerebellar granule cells differentiated in culture. A compositional study. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 16. pp. 11658-11665.
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