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Targeted proteomics to study mitochondrial biology

Wolters, J. C., Permentier, H., Bakker, B. & Bischoff, R., 30-Aug-2019, Mitochondria in health and in sickness. Urbani, A. & Babu, M. (eds.). Springer International Publishing AG, p. 101-117 17 p. (Advances in Experimental Medicine and Biology; vol. 1158).

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  • Targeted Proteomics to Study Mitochondrial Biology

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DOI

Targeted mass spectrometry in the selected or parallel reaction monitoring (SRM or PRM) mode is a widely used methodology to quantify proteins based on so-called signature or proteotypic peptides. SRM has the advantage of being able to quantify a range of proteins in a single analysis, for example, to measure the level of enzymes comprising a biochemical pathway. In this chapter, we will detail how to set up an SRM assay on the example of the mitochondrial protein succinate dehydrogenase [ubiquinone] flavoprotein subunit (mouse UniProt-code Q8K2B3). First, we will outline the in silico assay design including the choice of peptides based on a range of properties. We will further delineate different quantification strategies and introduce the reader to LC-MS assay development including the selection of the optimal peptide charge state and fragment ions as well as a discussion of the dynamic range of detection. The chapter will close with an application from the area of mitochondrial biology related to the quantification of a set of proteins isolated from mouse liver mitochondria in a study on mitochondrial respiratory flux decline in aging mouse muscle.
Original languageEnglish
Title of host publicationMitochondria in health and in sickness
EditorsA Urbani, M Babu
PublisherSpringer International Publishing AG
Chapter7
Pages101-117
Number of pages17
ISBN (Print)978-981-13-8366-3
Publication statusPublished - 30-Aug-2019

Publication series

NameAdvances in Experimental Medicine and Biology
PublisherSpringer
Volume1158
ISSN (Print)0065-2598
ISSN (Electronic)2214-8019

    Keywords

  • Targeted proteomics, Mitochondria, Mass spectrometry, Protein quantification, ABSOLUTE QUANTIFICATION, MASS-SPECTROMETRY, PROTEINS, MOUSE, IDENTIFICATION, IMMUNOASSAYS

ID: 95170952