Targeting mitochondria with small molecules: the preparation of MitoB and MitoP as exomarkers of mitochondrial hydrogen peroxide.

TitleTargeting mitochondria with small molecules: the preparation of MitoB and MitoP as exomarkers of mitochondrial hydrogen peroxide.
Publication TypeJournal Article
Year of Publication2015
AuthorsCairns, AG, McQuaker, SJ, Murphy, MP, Hartley, RC
JournalMethods Mol Biol
Volume1265
Pagination25-50
Date Published2015
ISSN1940-6029
KeywordsDrug Discovery, Hydrogen Peroxide, Mitochondria, Organophosphorus Compounds, Oxidation-Reduction, Phenols, Reactive Oxygen Species
Abstract

Small molecules can be physicochemically targeted to mitochondria using the lipophilic alkyltriphenylphosphonium (TPP) group. Once in the mitochondria the TPP-conjugate can detect or influence processes within the mitochondrial matrix directly. Alternatively, the conjugate can behave as a prodrug, which is activated by release from the TPP group either using an internal or external instruction. Small molecules can be designed that can be used in any cell line, tissue or whole organism, allow temporal control, and be applied in a reversible dose-dependent fashion. An example is the detection and quantification of hydrogen peroxide in mitochondria of whole living organisms by MitoB. Hydrogen peroxide produced within the mitochondrial matrix is involved in signalling and implicated in the oxidative damage associated with aging and a wide range of age-associated conditions including cardiovascular disease, neurodegeneration, and cancer. MitoB accumulates in mitochondria and is converted into the exomarker, MitoP, by hydrogen peroxide in the mitochondrial matrix. The hydrogen peroxide concentration is determined from the ratio of MitoP to MitoB after a period of incubation, and this ratio is determined by mass spectrometry using d15-MitoP and d15-MitoB as standard. Here we describe the synthesis of MitoB and MitoP and the deuterated standards necessary for this method of quantification.

DOI10.1007/978-1-4939-2288-8_3
Alternate JournalMethods Mol. Biol.
Citation Key10.1007/978-1-4939-2288-8_3
PubMed ID25634265
Grant ListMC_U105663142 / / Medical Research Council / United Kingdom