Immunoseparation of Prion protein‐enriched domains from other detergent‐resistant membrane fractions, isolated from neuronal cells

FEBS Letters

Botto, Laura²  Palestini, Paola²  Masserini, Massimo²  Cassetti, Arianna¹ 

[1] Consorzio MIA, Department of Neuroscience, University of Milano-Bicocca, Via Cadore 48, 20052 Monza, Italy;Department of Experimental, Environmental Medicine and Biotechnologies (DIMESAB), Medical School, University of Milano-Bicocca, Via Cadore 48, 20052 Monza, Italy

关键词: Neuron;    Lipid raft;    Prion protein;    PKC;    GAP43;    CGCs;    cerebellar granule cells;    DRM;    detergent-resistant membrane fraction;    MES;    2-[N-morpholino]ethansulfonic acid;    PMSF;    phenylmethylsulfonyl fluoride;   

DOI  :  10.1016/S0014-5793(03)01463-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 4°C, 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.

【 授权许可】

Unknown