Unknown

Dataset Information

0

Reductive amination of α-Ketoglutarate in metabolite extracts results in glutamate overestimation.


ABSTRACT: Artifacts due to metabolite extraction, derivatization, and detection techniques can result in aberrant observations that are not accurate representations of actual cell metabolism. Here, we show that α-ketoglutarate (α-KG) is reductively aminated to glutamate in methanol:water metabolite extracts, which introduces an artifact into metabolomics studies. We also identify pyridoxamine and urea as amine donors for α-KG to produce glutamate in methanol:water buffer in vitro, and we demonstrate that the addition of ninhydrin to the methanol:water buffer suppresses the reductive amination of α-KG to glutamate in vitro and in metabolite extracts. Finally, we calculate that glutamate levels have been overestimated by 10-50%, depending on cell line, due to α-KG reductive amination. These findings suggest that precautions to account for α-KG reductive amination should be taken for the accurate quantification of glutamate in metabolomics studies.

SUBMITTER: Wiese EK 

PROVIDER: S-EPMC7295581 | biostudies-literature |

REPOSITORIES: biostudies-literature

Similar Datasets

| S-EPMC1179207 | biostudies-other
| S-EPMC8253819 | biostudies-literature
| S-EPMC2819055 | biostudies-literature
| S-EPMC4057960 | biostudies-literature
| S-EPMC5925808 | biostudies-literature
| S-EPMC8048798 | biostudies-literature
| S-EPMC8508957 | biostudies-literature
| S-EPMC9864054 | biostudies-literature
| S-EPMC6594794 | biostudies-literature
| S-EPMC6698183 | biostudies-literature