Advanced

Relative Quantification of Membrane Proteins in Wild-Type and Prion Protein (PrP)-Knockout Cerebellar Granule Neurons

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 (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:
author
organization
publishing date
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
external identifiers
  • wos:000300458300002
  • scopus:84856641537
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
2012-05-15 07:42:16
date last changed
2017-01-01 03:59:49
@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},
  keyword      = {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},
  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},
  volume       = {11},
  year         = {2012},
}