A mitochondria-targeted mass spectrometry probe to detect glyoxals: implications for diabetes.

TitleA mitochondria-targeted mass spectrometry probe to detect glyoxals: implications for diabetes.
Publication TypeJournal Article
Year of Publication2014
AuthorsPun, PBoon Li, Logan, A, Darley-Usmar, V, Chacko, B, Johnson, MS, Huang, GW, Rogatti, S, Prime, TA, Methner, C, Krieg, T, Fearnley, IM, Larsen, L, Larsen, DS, Menger, KE, Collins, Y, James, AM, Kumar, GDKishore, Hartley, RC, Smith, RAJ, Murphy, MP
JournalFree Radic Biol Med
Volume67
Pagination437-50
Date Published2014 Feb
ISSN1873-4596
KeywordsAnimals, Cattle, Cell Line, Chromatography, Liquid, Diabetes Mellitus, Type 1, Disease Models, Animal, Endothelial Cells, Glyoxal, Hyperglycemia, Mice, Mitochondria, Liver, Molecular Probes, Myoblasts, Organophosphorus Compounds, Oxidative Stress, Phenylenediamines, Pyruvaldehyde, Rats, Tandem Mass Spectrometry
Abstract

The glycation of protein and nucleic acids that occurs as a consequence of hyperglycemia disrupts cell function and contributes to many pathologies, including those associated with diabetes and aging. Intracellular glycation occurs after the generation of the reactive 1,2-dicarbonyls methylglyoxal and glyoxal, and disruption of mitochondrial function is associated with hyperglycemia. However, the contribution of these reactive dicarbonyls to mitochondrial damage in pathology is unclear owing to uncertainties about their levels within mitochondria in cells and in vivo. To address this we have developed a mitochondria-targeted reagent (MitoG) designed to assess the levels of mitochondrial dicarbonyls within cells. MitoG comprises a lipophilic triphenylphosphonium cationic function, which directs the molecules to mitochondria within cells, and an o-phenylenediamine moiety that reacts with dicarbonyls to give distinctive and stable products. The extent of accumulation of these diagnostic heterocyclic products can be readily and sensitively quantified by liquid chromatography-tandem mass spectrometry, enabling changes to be determined. Using the MitoG-based analysis we assessed the formation of methylglyoxal and glyoxal in response to hyperglycemia in cells in culture and in the Akita mouse model of diabetes in vivo. These findings indicated that the levels of methylglyoxal and glyoxal within mitochondria increase during hyperglycemia both in cells and in vivo, suggesting that they can contribute to the pathological mitochondrial dysfunction that occurs in diabetes and aging.

DOI10.1016/j.freeradbiomed.2013.11.025
Alternate JournalFree Radic. Biol. Med.
Citation Key10.1016/j.freeradbiomed.2013.11.025
PubMed ID24316194
PubMed Central IDPMC3978666
Grant ListBB/I012826/1 / / Biotechnology and Biological Sciences Research Council / United Kingdom
BB/I012923/1 / / Biotechnology and Biological Sciences Research Council / United Kingdom
DK075865 / DK / NIDDK NIH HHS / United States
MC_U105663142 / / Medical Research Council / United Kingdom
P30 DK079626 / DK / NIDDK NIH HHS / United States
R01 DK075865 / DK / NIDDK NIH HHS / United States
/ / Medical Research Council / United Kingdom