The uploaded metabolomic dataset contains liquid-chromatography-mass spectrometry (LC-MS) data associated to a publication of Bruna Martins Garcia et al. from the Lena Pernas laboratory. The title olf the article is: Glutamine sensing licenses cholesterol synthesis. This article is to be published 2024 in the EMBO Journal- Abstract of the article: The mevalonate pathway produces essential metabolites such as cholesterol. Although this pathway is negatively regulated by metabolic intermediates, little is known of the metabolites that positively regulate its activity. We found that the amino acid glutamine is required to activate the mevalonate pathway. Glutamine starvation inhibited cholesterol synthesis and blocked transcription of the mevalonate pathway—even in the presence of glutamine derivatives such as ammonia and a-ketoglutarate. We pinpointed this glutamine-dependent effect to a loss in the ER-to-Golgi trafficking of SCAP that licenses the activation of SREBP2, the major transcriptional regulator of cholesterol synthesis. Both enforced Golgi-to-ER retro-translocation and the expression of a nuclear SREBP2 rescued mevalonate pathway activity during glutamine starvation. In a cell model of impaired mitochondrial respiration in which glutamine uptake is enhanced, SREBP2 activation and cellular cholesterol were increased. Thus, the mevalonate pathway senses and is activated by glutamine at a previously uncharacterized step, and the modulation of glutamine synthesis may be a strategy to regulate cholesterol levels in pathophysiological conditions. The associated data in this repository is grouped according to the figures in the the above mentioned article. Each zip folder contains the LC-MS raw files and one or more Excel tables describing the parameters (retention time, observed molecular weight, detected error to expected molecular weight, signal-to-noise and the integrated raw values of the detected compounds. Material and Method utilized for the analysis of the diverse samples is available in the context of the above mentioned article.