A sensitive mass spectrometric assay for mitochondrial CoQ pool redox state in vivo

Nils Burger; Angela Logan; Tracy A Prime; Amin Mottahedin; Stuart T Caldwell; Thomas Krieg; Richard C Hartley; Andrew M James; Michael P Murphy

doi:10.1016/j.freeradbiomed.2019.11.028

A sensitive mass spectrometric assay for mitochondrial CoQ pool redox state in vivo

Free Radic Biol Med. 2020 Feb 1:147:37-47. doi: 10.1016/j.freeradbiomed.2019.11.028. Epub 2019 Dec 5.

Authors

Nils Burger¹, Angela Logan¹, Tracy A Prime¹, Amin Mottahedin², Stuart T Caldwell³, Thomas Krieg⁴, Richard C Hartley³, Andrew M James¹, Michael P Murphy⁵

Affiliations

¹ MRC Mitochondrial Biology Unit, University of Cambridge, Hills Road, Cambridge, CB2 0XY, UK.
² MRC Mitochondrial Biology Unit, University of Cambridge, Hills Road, Cambridge, CB2 0XY, UK; Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge, CB2 0QQ, UK; Department of Physiology, Institute of Neuroscience and Physiology, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden.
³ School of Chemistry, University of Glasgow, Glasgow, G12 8QQ, UK.
⁴ Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge, CB2 0QQ, UK.
⁵ MRC Mitochondrial Biology Unit, University of Cambridge, Hills Road, Cambridge, CB2 0XY, UK; Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge, CB2 0QQ, UK. Electronic address: mpm@mrc-mbu.cam.ac.uk.

PMID: 31811922
PMCID: PMC6975167
DOI: 10.1016/j.freeradbiomed.2019.11.028

Abstract

Coenzyme Q (CoQ) is an essential cofactor, primarily found in the mitochondrial inner membrane where it functions as an electron carrier in the respiratory chain, and as a lipophilic antioxidant. The redox state of the CoQ pool is the ratio of its oxidised (ubiquinone) and reduced (ubiquinol) forms, and is a key indicator of mitochondrial bioenergetic and antioxidant status. However, the role of CoQ redox state in vivo is poorly understood, because determining its value is technically challenging due to redox changes during isolation, extraction and analysis. To address these problems, we have developed a sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay that enables us to extract and analyse both the CoQ redox state and the magnitude of the CoQ pool with negligible changes to redox state from small amounts of tissue. This will enable the physiological and pathophysiological roles of the CoQ redox state to be investigated in vivo.

Keywords: CoQ(10); CoQ(9); Coenzyme Q; Mass spectrometry; Mitochondria; Oxidative stress; Redox state.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromatography, Liquid
Mitochondria / metabolism
Oxidation-Reduction
Tandem Mass Spectrometry*
Ubiquinone* / metabolism

Substances

Ubiquinone

Abstract

Publication types

MeSH terms

Substances

Grants and funding