Summary of Study ST002542

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001637. The data can be accessed directly via it's Project DOI: 10.21228/M83M7P This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST002542
Study Title	Methionine restriction constrains lipoylation and activates mitochondria for nitrogenic synthesis of amino acids (Part 2)
Study Summary	Methionine restriction (MR) provides metabolic benefits in many organisms. However, mechanisms underlying the MR-induced effect remain incompletely understood. Here, we show in the budding yeast S. cerevisiae that MR relays a signal of S-adenosylmethionine (SAM) deprivation to adapt bioenergetic mitochondria to nitrogenic anabolism. In particular, decreases in cellular SAM constrain lipoate metabolism and protein lipoylation required for the operation of the tricarboxylic acid (TCA) cycle in the mitochondria, leading to incomplete glucose oxidation with an exit of acetyl-CoA and alpha-ketoglutarate from the TCA cycle to the syntheses of amino acids, such as arginine and leucine. This mitochondrial SAM-induced response, namely mitoSIR, promotes cell fitness through the coordination of mitochondrial fuel metabolism with the nitrogenic synthesis of amino acids.
Institute	Life Sciences Institute, ZheJiang University
Last Name	Cunqi
First Name	Ye
Address	866 Yuhangtang Rd, Hangzhou 310058, P.R. China
Email	yecunqi@zju.edu.cn
Phone	15267181160
Submit Date	2023-04-05
Raw Data Available	Yes
Raw Data File Type(s)	wiff
Analysis Type Detail	LC-MS
Release Date	2023-04-21
Release Version	1

Select appropriate tab below to view additional metadata details:

Sample Preparation:

Sampleprep ID:	SP002648
Sampleprep Summary:	Briefly, equal OD units of cells were rapidly quenched to stop metabolism by addition into 4 volumes of quenching buffer containing 60% methanol and 10 mM Tricine, pH 7.4 that was pre-cooled to -40°C. 5 min after holding at -40°C, cells were spun at 5,000 g for 3 min at 0°C, washed with the same buffer, and then resuspended in 1 mL extraction buffer containing 75% ethanol and 0.5 mM Tricine, pH 7.4. Intracellular metabolites were extracted by incubating at 75°C for 3 min, followed by chilling on ice for 5 min. Samples were spun at 20,000 g for 1 min to pellet cell debris, and 0.9 mL of the supernatant was transferred to a new tube. After a second spin at 20,000 g for 10 min, 0.8 mL of the supernatant was transferred to a new tube. Metabolites in the extraction buffer were dried using a vacuum concentrator system (Labconco) and stored at -80°C until analysis. Dried metabolite extracts were resuspended in 60% acetonitrile for injection.

Sampleprep ID:

SP002648

Sampleprep Summary:

Briefly, equal OD units of cells were rapidly quenched to stop metabolism by addition into 4 volumes of quenching buffer containing 60% methanol and 10 mM Tricine, pH 7.4 that was pre-cooled to -40°C. 5 min after holding at -40°C, cells were spun at 5,000 g for 3 min at 0°C, washed with the same buffer, and then resuspended in 1 mL extraction buffer containing 75% ethanol and 0.5 mM Tricine, pH 7.4. Intracellular metabolites were extracted by incubating at 75°C for 3 min, followed by chilling on ice for 5 min. Samples were spun at 20,000 g for 1 min to pellet cell debris, and 0.9 mL of the supernatant was transferred to a new tube. After a second spin at 20,000 g for 10 min, 0.8 mL of the supernatant was transferred to a new tube. Metabolites in the extraction buffer were dried using a vacuum concentrator system (Labconco) and stored at -80°C until analysis. Dried metabolite extracts were resuspended in 60% acetonitrile for injection.