Relative Quantification of Membrane Proteins in Wild-Type and Prion Protein (PrP)-Knockout Cerebellar Granule Neurons
(2012) In Journal of Proteome Research 11(2). p.523-536- Abstract
- 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... (More)
- 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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/2517429
- author
- Stella, Roberto ; Cifani, Paolo LU ; Peggion, Caterina ; Hansson, Karin M LU ; Lazzari, Cristian ; Bendz, Maria ; Levander, Fredrik LU ; Sorgato, Maria Catia ; Bertoli, Alessandro and James, Peter LU
- organization
- publishing date
- 2012
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- tandem mass tags, multiple reaction monitoring, mass spectrometry, gene knockout, membrane proteins, Prion protein, PrP
- in
- Journal of Proteome Research
- volume
- 11
- issue
- 2
- pages
- 523 - 536
- publisher
- The American Chemical Society (ACS)
- external identifiers
-
- wos:000300458300002
- scopus:84856641537
- pmid:22023170
- ISSN
- 1535-3893
- DOI
- 10.1021/pr200759m
- language
- English
- LU publication?
- yes
- id
- 43fe710e-a4a0-42e9-b8e6-a9cf44608747 (old id 2517429)
- date added to LUP
- 2016-04-01 10:54:09
- date last changed
- 2024-01-07 03:59:40
@article{43fe710e-a4a0-42e9-b8e6-a9cf44608747, abstract = {{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.}}, author = {{Stella, Roberto and Cifani, Paolo and Peggion, Caterina and Hansson, Karin M and Lazzari, Cristian and Bendz, Maria and Levander, Fredrik and Sorgato, Maria Catia and Bertoli, Alessandro and James, Peter}}, issn = {{1535-3893}}, keywords = {{tandem mass tags; multiple reaction monitoring; mass spectrometry; gene knockout; membrane proteins; Prion protein; PrP}}, language = {{eng}}, number = {{2}}, pages = {{523--536}}, publisher = {{The American Chemical Society (ACS)}}, series = {{Journal of Proteome Research}}, title = {{Relative Quantification of Membrane Proteins in Wild-Type and Prion Protein (PrP)-Knockout Cerebellar Granule Neurons}}, url = {{http://dx.doi.org/10.1021/pr200759m}}, doi = {{10.1021/pr200759m}}, volume = {{11}}, year = {{2012}}, }