Assay for methylmalonyl coenzyme A mutase activity based on determination of succinyl coenzyme A by ultrahigh-performance liquid chromatography tandem mass spectrometry

Gotoh, Kana; Nakajima, Yoko; Tanaka, Go; Hotta, Yuji; Kataoka, Tomoya; Kawade, Yoshihiro; Sugiyama, Naruji; Ito, Tetsuya; Kimura, Kazunori; Maeda, Yasuhiro

doi:10.1007/s00216-015-8753-8

Cited by 6 publications

(9 citation statements)

References 16 publications

(17 reference statements)

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“…The same pattern was observed for all analysed acyl-CoAs, including the reference standards, independent of the structure of the acyl rest. Importantly, we have not observed any fragment ions resulting from the acyl rest in agreement with all previously published data [2,7,10,28], with the only exception being the decarboxylation of malonyl-CoA derivatives mentioned above. The next very important criterion is the mass accuracy of the ions [M+ H] + and [M+H-506.9952] + .…”

Section: Methods Comparisonsupporting

confidence: 93%

“…1b). In addition, the CoA monothioesters of malonic acid derivatives are distinguishable from those of other dicarboxylic acids by mass spectrometry as reported previously [11,28]. Namely, in the mass spectra of both malonyl-CoA derivatives, a characteristic decarboxylation fragment is observed, which is not present in the spectra of the succinyl-CoA derivatives (Fig.…”

Section: Liquid Chromatographysupporting

confidence: 76%

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Suspect screening and targeted analysis of acyl coenzyme A thioesters in bacterial cultures using a high-resolution tribrid mass spectrometer

et al. 2021

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Analysis of acyl coenzyme A thioesters (acyl-CoAs) is crucial in the investigation of a wide range of biochemical reactions and paves the way to fully understand the concerned metabolic pathways and their superimposed networks. We developed two methods for suspect screening of acyl-CoAs in bacterial cultures using a high-resolution Orbitrap Fusion tribrid mass spectrometer. The methods rely on specific fragmentation patterns of the target compounds, which originate from the coenzyme A moiety. They make use of the formation of the adenosine 3′,5′-diphosphate key fragment (m/z 428.0365) and the neutral loss of the adenosine 3′-phosphate-5′-diphosphate moiety (506.9952) as preselection criteria for the detection of acyl-CoAs. These characteristic ions are generated either by an optimised in-source fragmentation in a full scan Orbitrap measurement or by optimised HCD fragmentation. Additionally, five different filters are included in the design of method. Finally, data-dependent MS/MS experiments on specifically preselected precursor ions are performed. The utility of the methods is demonstrated by analysing cultures of the denitrifying betaproteobacterium “Aromatoleum” sp. strain HxN1 anaerobically grown with hexanoate. We detected 35 acyl-CoAs in total and identified 24 of them by comparison with reference standards, including all 9 acyl-CoA intermediates expected to occur in the degradation pathway of hexanoate. The identification of additional acyl-CoAs provides insight into further metabolic processes occurring in this bacterium. The sensitivity of the method described allows detecting acyl-CoAs present in biological samples in highly variable abundances.

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Section: Methods Comparisonsupporting

confidence: 93%

Section: Liquid Chromatographysupporting

confidence: 76%

Suspect screening and targeted analysis of acyl coenzyme A thioesters in bacterial cultures using a high-resolution tribrid mass spectrometer

et al. 2021

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“…Analysis of samples was performed on an Acquity ultra-performance LC system coupled to a Quattro Premier XE triple quadrupole MS using an Acquity UPLC BEH C18 1.7 m 2.1 × 100-mm column (Waters Corporation). This is a modified version of the method by Gotoh et al (2015). The LC operation used the mobile phases: 400 mM ammonium formate (A) and acetonitrile (B).…”

Section: Analysis Of Samplesmentioning

confidence: 99%

Hepatic metabolism of propionate relative to meals for cows in the postpartum period

Kennedy

Allen

2019

Journal of Dairy Science

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The objective of this research was to identify potential short-term metabolic bottlenecks of propionate metabolism in the liver of dairy cows in the postpartum (PP) period and how such bottlenecks are affected by feeding status. Propionate, produced primarily from the fermentation of starch, decreases dry matter intake for cows in the postpartum period, likely by stimulating oxidation of acetyl-CoA in the liver. In this study, 8 dairy cows [2 blocks of 4 cows each, 6.63 ± 1.19 (mean ± SD) days PP; body condition score of 2.84 ± 0.39] were administered a pulse dose of either 1.5 mol/500 mL of propionic acid (PA) or 500 mL of water (control; CON) to the rumen either 1 h before or 2 h after feeding in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. Liver tissue was sampled at −1, 10 and 20 min relative to dosing, and blood was sampled at −30, −20, −10, −1, 5, 10, 15, 20, 25, 30, and 60 min relative to dosing. We hypothesized that rapid propionate absorption results in bottlenecks as enzymes become saturated and cofactors require regeneration. The PA treatment increased plasma propionate and insulin concentrations rapidly, with peaks reached by 5 min regardless of feeding status and cleared from the plasma within 30 min of dosing. The PA treatment decreased plasma nonesterified fatty acid concentration over 30 min compared with CON before but not after feeding. The PA treatment decreased plasma β-hydroxybutyrate concentration and increased plasma lactate concentration compared with CON both before and after feeding. The PA treatment also increased hepatic pyruvate and lactate concentrations compared with CON. The PA treatment tended to increase hepatic isocitrate and fumarate concentrations but did not affect hepatic malate and oxaloacetate concentrations, suggesting that elevated mitochondrial NADH/NAD + may have slowed the isocitrate dehydrogenase and fumarase reactions. The PA treatment also increased succinate concentration compared with CON, suggesting that a bottleneck may be present at succinate dehydrogenase. The PA treatment tended to increase citrate concentration despite having no effects on acetyl-CoA or oxaloacetate concentrations. These results are in agreement with our hypothesis that rapid absorption of propionate from the rumen and extraction by the liver results in metabolic bottlenecks in the liver that may affect feeding behavior and dry matter intake in dairy cows in the PP period.

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“…MCM activity was analyzed using a previously described method based on ultrahigh-performance LC/MS/MS. [15] MCM was obtained from 5×10 5 lymphocytes in Tris-sulfate buffer (pH 7.5) lysed by sonic disruption, and then adenosylcobalamin was added to the resultant lysate. After this solution was warmed at 37°C for a few minutes, it was spiked with methylmalonyl-CoA and incubated at 37°C for 15 min.…”

Section: Analysis Of MCM Activitymentioning

confidence: 99%

Clinical Application of Liquid Chromatography Tandem Mass Spectrometry Using Dried Blood Spot as a More Rapid Method for Determination of Methylmalonic Acid, Propionylcarnitine, and Total Homocysteine

Iijima

Ishige

Kubota

2020

J. inborn errors metab. screen.

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Methylmalonic acidemia (MMA) should be diagnosed in early infancy and receive appropriate management promptly after the diagnosis to prevent severe complications leading to death. At present, a newborn screening (NBS) method using tandem mass spectrometry (MS/MS) identifies suspected patients with MMA by elevated propionylcarnitine. In addition, a liquid chromatography tandem mass spectrometry (LC/MS/MS) method using dried blood spot is effective to detect some metabolites as a second-tier test, and reduces the false-positive rate in NBS. However, these tests were only used in screening, and not applied as an examination for evaluating treatment. Herein, we describe a 57-day-old girl with MMA under treatment with cobalamin who had elevated urinary methylmalonic acid levels. We applied the LC/MS/MS method with a separation column to evaluate her cobalamin responsiveness, and discovered an insufficient cobalamin dose earlier than would have been possible using other methods. Based on the current data, this method seems to be applicable for the follow-up of the treatment of MMA patients. However, this should be confirmed with more experience with a larger number of cases and a wider spectrum of disorders.

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Assay for methylmalonyl coenzyme A mutase activity based on determination of succinyl coenzyme A by ultrahigh-performance liquid chromatography tandem mass spectrometry

Cited by 6 publications

References 16 publications

Suspect screening and targeted analysis of acyl coenzyme A thioesters in bacterial cultures using a high-resolution tribrid mass spectrometer

Suspect screening and targeted analysis of acyl coenzyme A thioesters in bacterial cultures using a high-resolution tribrid mass spectrometer

Hepatic metabolism of propionate relative to meals for cows in the postpartum period

Clinical Application of Liquid Chromatography Tandem Mass Spectrometry Using Dried Blood Spot as a More Rapid Method for Determination of Methylmalonic Acid, Propionylcarnitine, and Total Homocysteine

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