Du är här

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

Författare:
Publiceringsår: 2012
Språk: Engelska
Sidor: 523-536
Publikation/Tidskrift/Serie: Journal of Proteome Research
Volym: 11
Nummer: 2
Dokumenttyp: Artikel
Förlag: American Chemical Society

Sammanfattning

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.

Disputation

Nyckelord

  • Medicine and Health Sciences
  • tandem mass tags
  • multiple reaction monitoring
  • mass spectrometry
  • gene knockout
  • membrane proteins
  • Prion protein
  • PrP

Övriga

Published
Yes
  • ISSN: 1535-3893

Box 117, 221 00 LUND
Telefon 046-222 00 00 (växel)
Telefax 046-222 47 20
lu [at] lu [dot] se

LERU logotype U21 logotype

Fakturaadress: Box 188, 221 00 LUND
Organisationsnummer: 202100-3211
Om webbplatsen