Development of a chemical probe for identifying protein targets of α-oxoaldehydes

Sibbersen, Christian; Palmfeldt, Johan; Hansen, Jens Christian; Gregersen, Niels; Jørgensen, Karl Anker; Johannsen, Mogens

doi:10.1039/c3cc41099d

Cited by 36 publications

(36 citation statements)

References 32 publications

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“…The enriched binding site peptides were released and analyzed by nano-liquid chromatography-tandem mass spectrometry (nanoLC-MS/MS). In previous work, we demonstrated high concurrence between MG and alk-MG binding sites on model proteins (Sibbersen et al, 2013) and, although we assume similar behavior for 3-HHD and alk-3-HHD, the addition of the propargyl group could potentially influence labeling of sterically congested lysine residues. The analysis led to the identification of several sites of HTO-and HDMP-type modification on plasma proteins (Table S4), predominantly on serum albumin.…”

Section: -Hhd Binds and Modifies Proteinsmentioning

confidence: 52%

“…We observed a clear dose-and time-dependent labeling of multiple proteins within the plasma fraction but not within the cellular fraction (Figures 5B and S5D), strongly indicating that 3-HHD is metabolized to less-reactive species by the blood cells. Curiously, the labeling with alk-3-HHD exceeded that of an analogous alkMG (6) probe that we have previously reported (Sibbersen et al, 2013), presumably as the latter is subject to rapid glyoxalasedependent turnover in whole blood. However, side-by-side comparison of the reactivity toward human plasma proteins clearly demonstrated the higher reactivity of the MG probe ( Figures S5E and S5F).…”

Section: -Hhd Binds and Modifies Proteinsmentioning

confidence: 70%

“…Refer to the Key Resources Table. 3-HHD and its isotopes as well as alkMG were synthesized and purified as described previously (Alberg et al, 2009;Sibbersen et al, 2013). HTO and hexane-2,3,5-triol were synthesized and purified as described in Methods S1.…”

Section: Methods Details Chemicalsmentioning

confidence: 99%

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Ketone Body Acetoacetate Buffers Methylglyoxal via a Non-enzymatic Conversion during Diabetic and Dietary Ketosis

Salomon

Sibbersen

Hansen

et al. 2017

Cell Chemical Biology

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The α-oxoaldehyde methylglyoxal is a ubiquitous and highly reactive metabolite known to be involved in aging- and diabetes-related diseases. If not detoxified by the endogenous glyoxalase system, it exerts its detrimental effects primarily by reacting with biopolymers such as DNA and proteins. We now demonstrate that during ketosis, another metabolic route is operative via direct non-enzymatic aldol reaction between methylglyoxal and the ketone body acetoacetate, leading to 3-hydroxyhexane-2,5-dione. This novel metabolite is present at a concentration of 10%-20% of the methylglyoxal level in the blood of insulin-starved patients. By employing a metabolite-alkyne-tagging strategy it is clarified that 3-hydroxyhexane-2,5-dione is further metabolized to non-glycating species in human blood. The discovery represents a new direction within non-enzymatic metabolism and within the use of alkyne-tagging for metabolism studies and it revitalizes acetoacetate as a competent endogenous carbon nucleophile.

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Section: -Hhd Binds and Modifies Proteinsmentioning

confidence: 52%

Section: -Hhd Binds and Modifies Proteinsmentioning

confidence: 70%

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Ketone Body Acetoacetate Buffers Methylglyoxal via a Non-enzymatic Conversion during Diabetic and Dietary Ketosis

Salomon

Sibbersen

Hansen

et al. 2017

Cell Chemical Biology

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“…The reaction of MGO with nucleic acids induces DNA strand breaks and elevated mutation frequency (Lee et al, 1995;Murata-Kamiya et al, 2000). Reactions of MGO with proteins, preferentially directed to arginine residues, lead to structural and functional changes (Sibbersen et al, 2013). In humans, AGE accumulation increases with age and is associated with several pathologies, including diabetes as well as cardiovascular and neurodegenerative diseases (Goldin et al, 2006;Thornalley, 2008;Münch et al, 2012).…”

Section: Introductionmentioning

confidence: 99%

Defense against Reactive Carbonyl Species Involves at Least Three Subcellular Compartments Where Individual Components of the System Respond to Cellular Sugar Status

et al. 2017

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Methylglyoxal (MGO) and glyoxal (GO) are toxic reactive carbonyl species generated as by-products of glycolysis. The pre-emption pathway for detoxification of these products, the glyoxalase (GLX) system, involves two consecutive reactions catalyzed by GLXI and GLXII. In , the GLX system is encoded by three homologs of and three homologs of , from which several predicted GLXI and GLXII isoforms can be derived through alternative splicing. We identified the physiologically relevant splice forms using sequencing data and demonstrated that the resulting isoforms have different subcellular localizations. All three GLXI homologs are functional in vivo, as they complemented a yeast GLXI loss-of-function mutant. Efficient MGO and GO detoxification can be controlled by a switch in metal cofactor usage. MGO formation is closely connected to the flux through glycolysis and through the Calvin Benson cycle; accordingly, expression analysis indicated that GLXI is transcriptionally regulated by endogenous sugar levels. Analyses of Arabidopsis loss-of-function lines revealed that the elimination of toxic reactive carbonyl species during germination and seedling establishment depends on the activity of the cytosolic GLXI;3 isoform. The Arabidopsis GLX system involves the cytosol, chloroplasts, and mitochondria, which harbor individual components that might be used at specific developmental stages and respond differentially to cellular sugar status.

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“…The reagent 2-oxohex-5-ynal was prepared according to the previous literature [5]. The 100-mer single-stranded DNA oligonucleotides (SS-G), 5′-(CAG TGA AGT TGG CAG ACT GAG CCA GGT CCC ACA GAT GCA GTG ACC GGA GTC ATT GCC AAA CTC TGC AGG AGA GCA AGG GCT GTC TAT AGG TGG CAA GTC A)-3′, were custom-synthesized (GeneDesign, Japan).…”

Section: Methodsmentioning

confidence: 99%

Biotinylation of Deoxyguanosine at the Abasic Site in Double-Stranded Oligodeoxynucleotides

2016

Journal of Analytical Methods in Chemistry

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Biotinylation of deoxyguanosine at an abasic site in double-stranded oligodeoxynucleotides was studied. The biotinylation of deoxyguanosine is achieved by copper-catalyzed click reaction after the conjugation of the oligodeoxynucleotide with 2-oxohex-5-ynal. The biotinylation enables visualization of the biotinylated oligodeoxynucleotides by chemiluminescence on a nylon membrane. In order to investigate the biotinylated site, the biotinylated oligodeoxynucleotides were amplified by the DNA polymerase chain reaction. Replacement of guanine opposing the abasic site with adenine generated by the activity of the terminal deoxynucleotidyl transferase of DNA polymerase was detected by DNA sequencing analysis and restriction endonuclease digestion. This study suggests that 2-oxohex-5-ynal may be useful for the detection of the unpaired deoxyguanosine endogenously generated at abasic sites in genomic DNA.

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Development of a chemical probe for identifying protein targets of α-oxoaldehydes

Abstract: The development of a chemical probe for identifying the protein targets of reactive electrophilic α-oxoaldehydes such as methylglyoxal is presented. The probe is evaluated against methylglyoxal using human serum albumin as well as using living cells and lysates.

Cited by 36 publications

References 32 publications

Ketone Body Acetoacetate Buffers Methylglyoxal via a Non-enzymatic Conversion during Diabetic and Dietary Ketosis

Ketone Body Acetoacetate Buffers Methylglyoxal via a Non-enzymatic Conversion during Diabetic and Dietary Ketosis

Defense against Reactive Carbonyl Species Involves at Least Three Subcellular Compartments Where Individual Components of the System Respond to Cellular Sugar Status

Biotinylation of Deoxyguanosine at the Abasic Site in Double-Stranded Oligodeoxynucleotides

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