Application of targeted mass spectrometry for the quantification of sirtuins in the central nervous system

T. Jayasena, A. Poljak, N. Braidy, L. Zhong, B. Rowlands, J. Muenchhoff, R. Grant, G. Smythe, C. Teo, M. Raftery, P. Sachdev*

*Corresponding author for this work

Research output: Contribution to journalArticle

9 Citations (Scopus)
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Abstract

Sirtuin proteins have a variety of intracellular targets, thereby regulating multiple biological pathways including neurodegeneration. However, relatively little is currently known about the role or expression of the 7 mammalian sirtuins in the central nervous system. Western blotting, PCR and ELISA are the main techniques currently used to measure sirtuin levels. To achieve sufficient sensitivity and selectivity in a multiplex-format, a targeted mass spectrometric assay was developed and validated for the quantification of all seven mammalian sirtuins (SIRT1-7). Quantification of all peptides was by multiple reaction monitoring (MRM) using three mass transitions per protein-specific peptide, two specific peptides for each sirtuin and a stable isotope labelled internal standard. The assay was applied to a variety of samples including cultured brain cells, mammalian brain tissue, CSF and plasma. All sirtuin peptides were detected in the human brain, with SIRT2 being the most abundant. Sirtuins were also detected in human CSF and plasma, and Guinea pig and mouse tissues. In conclusion, we have successfully applied MRM mass spectrometry for the detection and quantification of sirtuin proteins in the central nervous system, paving the way for more quantitative and functional studies.

Original languageEnglish
Article number35391
Pages (from-to)1-11
Number of pages11
JournalScientific Reports
Volume6
DOIs
Publication statusPublished - 20 Oct 2016
Externally publishedYes

Bibliographical note

Copyright the Author(s) 2016. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.

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    Jayasena, T., Poljak, A., Braidy, N., Zhong, L., Rowlands, B., Muenchhoff, J., ... Sachdev, P. (2016). Application of targeted mass spectrometry for the quantification of sirtuins in the central nervous system. Scientific Reports, 6, 1-11. [35391]. https://doi.org/10.1038/srep35391