The Comprehensive Native Interactome of a Fully Functional Tagged Prion Protein
Mertz, Kirsten D.
Calella, Anna Maria
- Journal Article
Rights / licenseCreative Commons Attribution 3.0 Unported
The enumeration of the interaction partners of the cellular prion protein, PrPC, may help clarifying its elusive molecular function. Here we added a carboxy proximal myc epitope tag to PrPC. When expressed in transgenic mice, PrPmyc carried a GPI anchor, was targeted to lipid rafts, and was glycosylated similarly to PrPC. PrPmyc antagonized the toxicity of truncated PrP, restored prion infectibility of PrPC-deficient mice, and was physically incorporated into PrPSc aggregates, indicating that it possessed all functional characteristics of genuine PrPC. We then immunopurified myc epitope-containing protein complexes from PrPmyc transgenic mouse brains. Gentle differential elution with epitope-mimetic decapeptides, or a scrambled version thereof, yielded 96 specifically released proteins. Quantitative mass spectrometry with isotope-coded tags identified seven proteins which co-eluted equimolarly with PrPC and may represent component of a multiprotein complex. Selected PrPC interactors were validated using independent methods. Several of these proteins appear to exert functions in axomyelinic maintenance. Show more
Journal / seriesPLoS ONE
Pages / Article No.
PublisherPublic Library of Science
Organisational unit02207 - Functional Genomics Center Zürich / Functional Genomics Center Zürich
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