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Peter James

Peter James

Professor

Peter James

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

Author

  • Roberto Stella
  • Paolo Cifani
  • Caterina Peggion
  • Karin M Hansson
  • Cristian Lazzari
  • Maria Bendz
  • Fredrik Levander
  • Maria Catia Sorgato
  • Alessandro Bertoli
  • Peter James

Summary, in English

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.

Department/s

  • Department of Immunotechnology

Publishing year

2012

Language

English

Pages

523-536

Publication/Series

Journal of Proteome Research

Volume

11

Issue

2

Document type

Journal article

Publisher

The American Chemical Society (ACS)

Topic

  • Immunology in the medical area
  • Health Sciences

Keywords

  • tandem mass tags
  • multiple reaction monitoring
  • mass spectrometry
  • gene knockout
  • membrane proteins
  • Prion protein
  • PrP

Status

Published

ISBN/ISSN/Other

  • ISSN: 1535-3893