Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin
The cellular prion protein (PrP<sup>C</sup>) is renowned for its infectious conformational isoform PrP<sup>Sc</sup>, capable of templating subsequent conversions of healthy PrP<sup>C</sup>s and thus triggering the group of incurable diseases known as transmissible...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-12-01
|
| Series: | Membranes |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2077-0375/11/12/978 |
| _version_ | 1827671213991788544 |
|---|---|
| author | Divya Teja Dondapati Pradeep Reddy Cingaram Ferhan Ayaydin Antal Nyeste Andor Kanyó Ervin Welker Elfrieda Fodor |
| author_facet | Divya Teja Dondapati Pradeep Reddy Cingaram Ferhan Ayaydin Antal Nyeste Andor Kanyó Ervin Welker Elfrieda Fodor |
| author_sort | Divya Teja Dondapati |
| collection | DOAJ |
| description | The cellular prion protein (PrP<sup>C</sup>) is renowned for its infectious conformational isoform PrP<sup>Sc</sup>, capable of templating subsequent conversions of healthy PrP<sup>C</sup>s and thus triggering the group of incurable diseases known as transmissible spongiform encephalopathies. Besides this mechanism not being fully uncovered, the protein’s physiological role is also elusive. PrP<sup>C</sup> and its newest, less understood paralog Shadoo are glycosylphosphatidylinositol-anchored proteins highly expressed in the central nervous system. While they share some attributes and neuroprotective actions, opposing roles have also been reported for the two; however, the amount of data about their exact functions is lacking. Protein–protein interactions and membrane microdomain localizations are key determinants of protein function. Accurate identification of these functions for a membrane protein, however, can become biased due to interactions occurring during sample processing. To avoid such artifacts, we apply a non-detergent-based membrane-fractionation approach to study the prion protein and Shadoo. We show that the two proteins occupy similarly raft and non-raft membrane fractions when expressed in N2a cells and that both proteins pull down the chaperone calnexin in both rafts and non-rafts. These indicate their possible binding to calnexin in both types of membrane domains, which might be a necessary requisite to aid the inherently unstable native conformation during their lifetime. |
| first_indexed | 2024-03-10T03:34:58Z |
| format | Article |
| id | doaj.art-2bf6305b1b1b4ff5a4a96ab862681d56 |
| institution | Directory Open Access Journal |
| issn | 2077-0375 |
| language | English |
| last_indexed | 2024-03-10T03:34:58Z |
| publishDate | 2021-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Membranes |
| spelling | doaj.art-2bf6305b1b1b4ff5a4a96ab862681d562023-11-23T09:30:44ZengMDPI AGMembranes2077-03752021-12-01111297810.3390/membranes11120978Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with CalnexinDivya Teja Dondapati0Pradeep Reddy Cingaram1Ferhan Ayaydin2Antal Nyeste3Andor Kanyó4Ervin Welker5Elfrieda Fodor6Institute of Biochemistry, Biological Research Centre, 6726 Szeged, HungaryInstitute of Biochemistry, Biological Research Centre, 6726 Szeged, HungaryHungarian Centre of Excellence for Molecular Medicine (HCEMM) Nonprofit Ltd., 6723 Szeged, HungaryProteoscientia Ltd., 3066 Cserhátszentiván, HungaryBiospirál-2006. Ltd., 6726 Szeged, HungaryInstitute of Biochemistry, Biological Research Centre, 6726 Szeged, HungaryInstitute of Biochemistry, Biological Research Centre, 6726 Szeged, HungaryThe cellular prion protein (PrP<sup>C</sup>) is renowned for its infectious conformational isoform PrP<sup>Sc</sup>, capable of templating subsequent conversions of healthy PrP<sup>C</sup>s and thus triggering the group of incurable diseases known as transmissible spongiform encephalopathies. Besides this mechanism not being fully uncovered, the protein’s physiological role is also elusive. PrP<sup>C</sup> and its newest, less understood paralog Shadoo are glycosylphosphatidylinositol-anchored proteins highly expressed in the central nervous system. While they share some attributes and neuroprotective actions, opposing roles have also been reported for the two; however, the amount of data about their exact functions is lacking. Protein–protein interactions and membrane microdomain localizations are key determinants of protein function. Accurate identification of these functions for a membrane protein, however, can become biased due to interactions occurring during sample processing. To avoid such artifacts, we apply a non-detergent-based membrane-fractionation approach to study the prion protein and Shadoo. We show that the two proteins occupy similarly raft and non-raft membrane fractions when expressed in N2a cells and that both proteins pull down the chaperone calnexin in both rafts and non-rafts. These indicate their possible binding to calnexin in both types of membrane domains, which might be a necessary requisite to aid the inherently unstable native conformation during their lifetime.https://www.mdpi.com/2077-0375/11/12/978lipid raftsGPI-anchored proteinsendoplasmic reticulumdetergent-free raft isolation |
| spellingShingle | Divya Teja Dondapati Pradeep Reddy Cingaram Ferhan Ayaydin Antal Nyeste Andor Kanyó Ervin Welker Elfrieda Fodor Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin Membranes lipid rafts GPI-anchored proteins endoplasmic reticulum detergent-free raft isolation |
| title | Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin |
| title_full | Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin |
| title_fullStr | Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin |
| title_full_unstemmed | Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin |
| title_short | Membrane Domain Localization and Interaction of the Prion-Family Proteins, Prion and Shadoo with Calnexin |
| title_sort | membrane domain localization and interaction of the prion family proteins prion and shadoo with calnexin |
| topic | lipid rafts GPI-anchored proteins endoplasmic reticulum detergent-free raft isolation |
| url | https://www.mdpi.com/2077-0375/11/12/978 |
| work_keys_str_mv | AT divyatejadondapati membranedomainlocalizationandinteractionoftheprionfamilyproteinsprionandshadoowithcalnexin AT pradeepreddycingaram membranedomainlocalizationandinteractionoftheprionfamilyproteinsprionandshadoowithcalnexin AT ferhanayaydin membranedomainlocalizationandinteractionoftheprionfamilyproteinsprionandshadoowithcalnexin AT antalnyeste membranedomainlocalizationandinteractionoftheprionfamilyproteinsprionandshadoowithcalnexin AT andorkanyo membranedomainlocalizationandinteractionoftheprionfamilyproteinsprionandshadoowithcalnexin AT ervinwelker membranedomainlocalizationandinteractionoftheprionfamilyproteinsprionandshadoowithcalnexin AT elfriedafodor membranedomainlocalizationandinteractionoftheprionfamilyproteinsprionandshadoowithcalnexin |