Internalization of mammalian fluorescent cellular prion protein and N-terminal deletion mutants in living cells
| Autor: | Vilma R. Martins, Ricardo R. Brentani, Kil Sun Lee, Marco A. M. Prado, Ana C. Magalhães, Silvio M. Zanata |
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| Rok vydání: | 2008 |
| Předmět: |
Mutation
animal diseases media_common.quotation_subject Mutant Cell Biology Endocytosis medicine.disease_cause Biochemistry nervous system diseases Green fluorescent protein Cell biology Cellular and Molecular Neuroscience medicine.anatomical_structure Cell culture mental disorders medicine Internalization Intracellular media_common |
| Zdroj: | Journal of Neurochemistry. 79:79-87 |
| ISSN: | 0022-3042 |
| Popis: | The cellular prion protein (PrPc) is a glycosylphosphatidylinositol (GPI)-anchored plasma membrane protein whose conformational altered forms (PrPsc) are known to cause neurodegenerative diseases in mammals. In order to investigate the intracellular traffic of mammalian PrPc in living cells, we have generated a green fluorescent protein (GFP) tagged version of PrPc. The recombinant protein was properly anchored at the cell surface and its distribution pattern was similar to that of the endogenous PrPc, with labeling at the plasma membrane and in an intracellular perinuclear compartment. Comparison of the steady-state distribution of GFP-PrPc and two N-terminal deletion mutants (Δ32-121 and Δ32-134), that cause neurological symptoms when expressed in PrP knockout mice, was carried out. The mutant proteins accumulated in the plasma membrane at the expense of decreased labeling in the perinuclear region when compared with GFP-PrPc. In addition, GFP-PrPc, but not the two mutants, internalized from the plasma membrane in response to Cu2+ treatment and accumulated at a perinuclear region in SN56 cells. Our data suggest that GFP-PrPc can be used to follow constitutive and induced PrPc traffic in living cells. |
| Databáze: | OpenAIRE |
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