GONUTS has been updated to MW1.31 Most things seem to be working but be sure to report problems.
PMID:16294306
| Citation |
Strom, A, Diecke, S, Hunsmann, G and Stuke, AW (2006) Identification of prion protein binding proteins by combined use of far-Western immunoblotting, two dimensional gel electrophoresis and mass spectrometry. Proteomics 6:26-34 |
|---|---|
| Abstract |
The cellular prion protein (PrP(C)), a highly conserved glycoprotein predominantly expressed by neuronal cells, can convert into an abnormal isoform (PrP(Sc)) and provoke a transmissible spongiform encephalopathy. In spite of many studies, the physiological function of PrP(C) remains unknown. Recent findings suggest that PrP(C) is a multifunctional protein participating in several cellular processes. Using recombinant human PrP as a probe, we performed far-Western immunoblotting (protein overlay assay) to detect cellular PrP(C) interactors. Brain extracts of wild-type and PrP knockout mice were screened by far-Western immunoblotting for PrP-specific interactions. Subsequently, putative ligands were isolated by 2-DE and identified by MALDI-TOF MS, enabling identification of heterogeneous nuclear ribonucleoprotein A2/B1 and aldolase C as novel interaction partners of PrP(C). These data provide the first evidence of a molecule indicating a mechanism for the predicted involvement of PrP(C) in nucleic acid metabolisms. In summary, we have shown the successful combination of 2-DE with far-Western immunoblotting and MALDI-TOF MS for identification of new cellular binding partners of a known protein. Especially the application of this technique to investigate other neurodegenerative diseases is promising. |
| Links |
PubMed Online version:10.1002/pmic.200500066 |
| Keywords |
Animals; Blotting, Far-Western/methods; Brain/metabolism; Cytosol/metabolism; Electrophoresis, Gel, Two-Dimensional/methods; Heterogeneous-Nuclear Ribonucleoprotein Group A-B/metabolism; Immunoprecipitation; Membrane Proteins/metabolism; Mice; Nerve Tissue Proteins/metabolism; Prions/metabolism; Protein Binding; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO ID | GO term name | Evidence Code | with/from | Aspect | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| edit table |
See also
References
See Help:References for how to manage references in GONUTS.
