title: A mitochondria-targeted mass spectrometry probe to detect glyoxals: implications for diabetes.
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.
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.
authors: Pamela Boon Li
authors: PB
authors: Pun
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.
authors: Angela
authors: A
authors: Logan
authors: Department of Pathology, Centre for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors: Victor
authors: V
authors: Darley-Usmar
authors: Department of Pathology, Centre for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors: Balu
authors: B
authors: Chacko
authors: Department of Pathology, Centre for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors: Michelle S
authors: MS
authors: Johnson
authors: Department of Pathology, Centre for Free Radical Biology, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
authors: Guang W
authors: GW
authors: Huang
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.
authors: Sebastian
authors: S
authors: Rogatti
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.
authors: Tracy A
authors: TA
authors: Prime
authors: Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge CB2 2QQ, UK.
authors: Carmen
authors: C
authors: Methner
authors: Department of Medicine, University of Cambridge, Addenbrooke's Hospital, Cambridge CB2 2QQ, UK.
authors: Thomas
authors: T
authors: Krieg
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.
authors: Ian M
authors: IM
authors: Fearnley
authors: Department of Chemistry, University of Otago, Dunedin, New Zealand.
authors: Lesley
authors: L
authors: Larsen
authors: Department of Chemistry, University of Otago, Dunedin, New Zealand.
authors: David S
authors: DS
authors: Larsen
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.
authors: Katja E
authors: KE
authors: Menger
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.
authors: Yvonne
authors: Y
authors: Collins
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK.
authors: Andrew M
authors: AM
authors: James
authors: Centre for the Chemical Research of Ageing, WestCHEM School of Chemistry, University of Glasgow, Glasgow G12 8QQ, UK.
authors: G D Kishore
authors: GD
authors: Kumar
authors: Centre for the Chemical Research of Ageing, WestCHEM School of Chemistry, University of Glasgow, Glasgow G12 8QQ, UK.
authors: Richard C
authors: RC
authors: Hartley
authors: Department of Chemistry, University of Otago, Dunedin, New Zealand.
authors: Robin A J
authors: RA
authors: Smith
authors: MRC Mitochondrial Biology Unit, Wellcome Trust/MRC Building, Cambridge CB2 0XY, UK. Electronic address: mpm@mrc-mbu.cam.ac.uk.
authors: Michael P
authors: MP
authors: Murphy
mesh_terms: D000818:Animals; D002417:Cattle; D002460:Cell Line; D002853:Chromatography, Liquid; D003922:Diabetes Mellitus, Type 1 / Q000175:diagnosis / Q000378:metabolism* / Q000473:pathology; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology; D006037:Glyoxal / Q000032:analysis* / Q000378:metabolism; D006943:Hyperglycemia / Q000175:diagnosis / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D008930:Mitochondria, Liver / Q000378:metabolism* / Q000473:pathology; D015335:Molecular Probes / Q000138:chemical synthesis*; D032446:Myoblasts / Q000378:metabolism / Q000473:pathology; D009943:Organophosphorus Compounds / Q000737:chemistry; D018384:Oxidative Stress; D010655:Phenylenediamines / Q000737:chemistry; D011765:Pyruvaldehyde / Q000032:analysis* / Q000378:metabolism; D051381:Rats; D053719:Tandem Mass Spectrometry
chemical_list: D015335:Molecular Probes; D009943:Organophosphorus Compounds; D010655:Phenylenediamines; D006037:Glyoxal; D011765:Pyruvaldehyde
keywords: Exomarker; Free radicals; Glyoxal; Hyperglycemia; Methylglyoxal; MitoG; Mitochondria