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

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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.

Cifani, PaoloLund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)immu-pci (författare)
Peggion, Caterina (författare)
Hansson, Karin MLund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)klke-kh0 (författare)
Lazzari, Cristian (författare)
Bendz, MariaStockholms universitet,Institutionen för biokemi och biofysik(Swepub:su)mbend (författare)
Levander, FredrikLund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)tmb-fle (författare)
Sorgato, Maria Catia (författare)
Bertoli, Alessandro (författare)
James, PeterLund University,Lunds universitet,Institutionen för immunteknologi,Institutioner vid LTH,Lunds Tekniska Högskola,Department of Immunotechnology,Departments at LTH,Faculty of Engineering, LTH(Swepub:lu)elma-pja (författare)
Institutionen för immunteknologiInstitutioner vid LTH (creator_code:org_t)

Ingår i:Journal of Proteome Research: American Chemical Society (ACS)11:2, s. 523-5361535-38931535-3907

Av författaren/redakt...: Stella, Roberto; Cifani, Paolo; Peggion, Caterin ...; Hansson, Karin M; Lazzari, Cristia ...; Bendz, Maria; visa fler...; Levander, Fredri ...; Sorgato, Maria C ...; Bertoli, Alessan ...; James, Peter; visa färre...

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MEDICIN OCH HÄLSOVETENSKAP: MEDICIN OCH HÄLS ...; och Medicinska och f ...; och Immunologi inom ...

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