Mitochondrial damage in renal epithelial cells is potentiated by protein exposure in propionic aciduria

Schumann, Anke; Belche, Véronique; Schaller, K; Grünert, Sarah C.; Kaech, Andres; Baumgartner, Matthias R.; Kölker, Stefan; Hannibal, Luciana; Spiekerkoetter, Ute

doi:10.1002/jimd.12419

Cited by 8 publications

(2 citation statements)

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“…On the day of the experiment, urine samples were thawed, vortexed, and diluted 1:25 with water prior to sample preparation (see Section 4.5 ) for SAM, SAH, and creatinine determination. Human renal epithelial cells from proximal tubules and their corresponding conditioned culture media utilised in this study were grown, harvested, and stored as described in our previous work [ 58 ].…”

Section: Methodsmentioning

confidence: 99%

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Analysis of S-Adenosylmethionine and S-Adenosylhomocysteine: Method Optimisation and Profiling in Healthy Adults upon Short-Term Dietary Intervention

et al. 2022

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S-adenosylmethionine (SAM) is essential for methyl transfer reactions. All SAM is produced de novo via the methionine cycle. The demethylation of SAM produces S-adenosylhomocysteine (SAH), an inhibitor of methyltransferases and the precursor of homocysteine (Hcy). The measurement of SAM and SAH in plasma has value in the diagnosis of inborn errors of metabolism (IEM) and in research to assess methyl group homeostasis. The determination of SAM and SAH is complicated by the instability of SAM under neutral and alkaline conditions and the naturally low concentration of both SAM and SAH in plasma (nM range). Herein, we describe an optimised LC-MS/MS method for the determination of SAM and SAH in plasma, urine, and cells. The method is based on isotopic dilution and employs 20 µL of plasma or urine, or 500,000 cells, and has an instrumental running time of 5 min. The reference ranges for plasma SAM and SAH in a cohort of 33 healthy individuals (age: 19–60 years old; mean ± 2 SD) were 120 ± 36 nM and 21.5 ± 6.5 nM, respectively, in accordance with independent studies and diagnostic determinations. The method detected abnormal concentrations of SAM and SAH in patients with inborn errors of methyl group metabolism. Plasma and urinary SAM and SAH concentrations were determined for the first time in a randomised controlled trial of 53 healthy adult omnivores (age: 18–60 years old), before and after a 4 week intervention with a vegan or meat-rich diet, and revealed preserved variations of both metabolites and the SAM/SAH index.

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Section: Methodsmentioning

confidence: 99%

“…Cell pellets were lysed with ice-cold PBS supplemented with a 1% protease inhibitor cocktail as described [ 58 ]. Cells were disrupted by freeze-thawing and homogenisation with a cordless pestle motor and disposable pellet mixers (VWR Nr.…”

Section: Methodsmentioning

confidence: 99%