Mass spectrometry quantification of clusterin in the human brain

The multifunctional glycoprotein clusterin has been associated with late-onset Alzheimer's disease (AD). Further investigation to define the role of clusterin in AD phenotypes would be aided by the development of techniques to quantify level, potential post-translational modifications, and isof...

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Bibliographic Details
Published in:Molecular neurodegeneration 2012-08, Vol.7 (1), p.41-41, Article 41
Main Authors: Chen, Junjun, Wang, Meiyao, Turko, Illarion V
Format: Article
Language:English
Subjects:
Advertising executives
Alzheimer Disease - metabolism
Alzheimer's disease
Amino acids
Analysis
Autopsy
Brain
Brain - metabolism
Brain Chemistry
Clusterin
Clusterin - analysis
Clusterin - metabolism
Cortex (frontal)
Cortex (temporal)
Glycoproteins
Human brain
Humans
Mass spectrometry
Mass Spectrometry - methods
Mass spectroscopy
Methodology
Multiple reaction monitoring
Neurodegenerative diseases
Peptides
Phosphorylation
Post-translation
Protein Processing, Post-Translational - physiology
Proteins
QconCAT
Sodium lauryl sulfate
Translation
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Summary:The multifunctional glycoprotein clusterin has been associated with late-onset Alzheimer's disease (AD). Further investigation to define the role of clusterin in AD phenotypes would be aided by the development of techniques to quantify level, potential post-translational modifications, and isoforms of clusterin. We have developed a quantitative technique based on multiple reaction monitoring (MRM) mass spectrometry to measure clusterin in human postmortem brain tissues. A stable isotope-labeled concatenated peptide (QconCAT) bearing selected peptides from clusterin was expressed with an in vitro translation system and purified. This clusterin QconCAT was validated for use as an internal standard for clusterin quantification using MRM mass spectrometry. Measurements were performed on the human postmortem frontal and temporal cortex from control and severe AD cases. During brain tissues processing, 1% SDS was used in the homogenization buffer to preserve potential post-translational modifications of clusterin. However, MRM quantifications in the brain did not suggest phosphorylation of Thr(393), Ser(394), and Ser(396) residues reported for clusterin in serum. MRM quantifications in the frontal cortex demonstrated significantly higher (P 
ISSN:1750-1326
1750-1326
DOI:10.1186/1750-1326-7-41