Relative Quantification of Membrane Proteins in Wild-Type and Prion Protein (PrP)-Knockout Cerebellar Granule Neurons
2012 (English)In: Journal of Proteome Research, ISSN 1535-3893, E-ISSN 1535-3907, Vol. 11, no 2, 523-536 p.Article in journal (Refereed) Published
Approximately 25% of eukaryotic proteins possessing homology to at least two trans membrane domains are predicted to be embedded in biological membranes. Nevertheless, this group of proteins is not usually well represented in proteome-wide experiments due to their refractory nature. Here we present a quantitative mass spectrometry-based comparison of membrane protein expression in cerebellar granule neurons grown in primary culture that were isolated from wild-type mice and mice lacking the cellular prion protein. This protein is a cell-surface glycoprotein that is mainly expressed in the central nervous system and is involved in several neurodegenerative disorders, though its physiological role is unclear. We used a low specificity enzyme a-chymotrypsin to digest membrane proteins preparations that had been separated by SDS-PAGE. The resulting peptides were labeled with tandem mass tags and analyzed by MS. The differentially expressed proteins identified using this approach were further analyzed by multiple reaction monitoring to confirm the expression level changes.
Place, publisher, year, edition, pages
2012. Vol. 11, no 2, 523-536 p.
Prion protein, PrP, gene knockout, membrane proteins, mass spectrometry, multiple reaction monitoring, tandem mass tags
Biophysics Biochemistry and Molecular Biology
IdentifiersURN: urn:nbn:se:su:diva-76344DOI: 10.1021/pr200759mISI: 000300458300002OAI: oai:DiVA.org:su-76344DiVA: diva2:526398