The possibility of coexistence of different subtypes of membrane lipid rafts has been investigated in cerebellar granule cells, by submitting detergent-resistant membrane fractions to immunoprecipitation. Among the proteins and lipids present in detergent-resistant fractions, almost all Prion protein, GAP43 and PKC were present in the immunoprecipitate obtained with anti-GAP43 or anti-Prion protein antibody at 4degreesC, together with a small fraction of cholesterol and sphingolipids, suggesting that they belong to a distinct subset of membranes. On the contrary, all Fyn and almost all MARCKS remained in the supernatant. Fluorescence microscopy experiments showed that Fyn and Prion protein were mostly not colocalized within a single neuron. Our results suggest that granule cells membranes contains different subtypes of detergent-resistant fractions, possibly deriving from different lipid rafts. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Botto, L., Masserini, M., Cassetti, A., Palestini, P. (2004). Immunoseparation of Prion protein-enriched domains from other detergent-resistant membrane fractions, isolated from neuronal cells. FEBS LETTERS, 557(1-3), 143-147 [10.1016/S0014-5793(03)01463-7].
Immunoseparation of Prion protein-enriched domains from other detergent-resistant membrane fractions, isolated from neuronal cells
BOTTO, LAURA MARIA;MASSERINI, MASSIMO ERNESTO;PALESTINI, PAOLA NOVERINA ADA
2004
Abstract
The possibility of coexistence of different subtypes of membrane lipid rafts has been investigated in cerebellar granule cells, by submitting detergent-resistant membrane fractions to immunoprecipitation. Among the proteins and lipids present in detergent-resistant fractions, almost all Prion protein, GAP43 and PKC were present in the immunoprecipitate obtained with anti-GAP43 or anti-Prion protein antibody at 4degreesC, together with a small fraction of cholesterol and sphingolipids, suggesting that they belong to a distinct subset of membranes. On the contrary, all Fyn and almost all MARCKS remained in the supernatant. Fluorescence microscopy experiments showed that Fyn and Prion protein were mostly not colocalized within a single neuron. Our results suggest that granule cells membranes contains different subtypes of detergent-resistant fractions, possibly deriving from different lipid rafts. (C) 2003 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
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