Mass spectrometry quantification of clusterin in the human brain

dc.contributor.authorChen, Junjun
dc.contributor.authorWang, Meiyao
dc.contributor.authorTurko, Illarion
dc.date.accessioned2013-01-07T19:55:30Z
dc.date.available2013-01-07T19:55:30Z
dc.date.issued2012-08-20
dc.description.abstractBackground: 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. Results: 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 < 0.01) level of clusterin in severe AD group (39.1 ± 9.1 pmol/mg tissue protein) in comparison to control group (25.4 ± 4.4 pmol/mg tissue protein). In the temporal cortex, the clusterin levels were not significantly different, 29.0 ± 7.9 pmol/mg tissue protein and 28.0 ± 8.4 pmol/mg tissue protein in control and severe AD groups, respectively. Conclusions: The proposed protocol is a universal quantitative technique to assess expression level of clusterin. It is expected that application of this protocol to quantification of various clusterin isoforms and potential post-translational modifications will be helpful in addressing the role of clusterin in AD.en_US
dc.description.urihttps://doi.org/10.1186/1750-1326-7-41
dc.identifier.citationChen, J., Wang, M. & Turko, I.V. Mass spectrometry quantification of clusterin in the human brain. Mol Neurodegeneration 7, 41 (2012).en_US
dc.identifier.urihttp://hdl.handle.net/1903/13344
dc.language.isoen_USen_US
dc.relation.isAvailableAtCollege of Computer, Mathematical & Physical Sciencesen_us
dc.relation.isAvailableAtDigital Repository at the University of Marylanden_us
dc.relation.isAvailableAtBiologyen_us
dc.relation.isAvailableAtUniversity of Maryland (College Park, MD)en_us
dc.subjectClusterinen_US
dc.subjectQconCATen_US
dc.subjectMultiple reaction monitoringen_US
dc.subjectHuman brainen_US
dc.subjectAlzheimer’s diseaseen_US
dc.titleMass spectrometry quantification of clusterin in the human brainen_US
dc.typeArticleen_US

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