Summary of Study ST001382

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 PR000946. The data can be accessed directly via it's Project DOI: 10.21228/M8DM63 This work is supported by NIH grant, U2C- DK119886.

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Study ID	ST001382
Study Title	Distinct metabolic states of a cell guide alternate fates of mutational buffering through altered proteostasis
Study Summary	Changes in metabolism can alter the cellular milieu; can this also change intracellular proteostasis? Since proteostasis can modulate mutational buffering, if change in metabolism has the ability to change proteostasis, arguably, it should also alter mutational buffering. Building on this, we find that altered cellular metabolic states in E. coli buffer distinct mutations. Buffered-mutants had folding problems in vivo and were differently chaperoned in different metabolic states. Notably, this assistance was dependent upon the metabolites and not on the increase in canonical chaperone machineries. Additionally, we were able to reconstitute the folding assistance afforded by metabolites in vitro and propose that changes in metabolite concentrations have the potential to alter proteostasis. Collectively, we unravel that the metabolite pools are bona fide members of proteostasis and aid in mutational buffering. Given the plasticity in cellular metabolism, we posit that metabolic alterations may play an important role in the positive or negative regulation of proteostasis.
Institute	CSIR-National Chemical Laboratory
Last Name	Shanmugam
First Name	Dhanasekaran
Address	Dr. Homi Bhabha Road, Pune, maharashtra, 411008, India
Email	d.shanmugam@ncl.res.in
Phone	2025902719
Submit Date	2020-05-14
Raw Data Available	Yes
Raw Data File Type(s)	mzXML
Analysis Type Detail	LC-MS
Release Date	2020-06-01
Release Version	1

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Sample Preparation:

Sampleprep ID:	SP001464
Sampleprep Summary:	Cell pellets were extracted with 200ul of chilled extraction solvent (80% MetOH in MS grade water containing 1ng/µl PIPES and U13C-U15N-glutamine as internal standard) was added to the pellet, mixed and incubated in ice for 5 minutes for quenching metabolism. This is followed by sonication in water bath for 15 min at 4°C with intermittent vortexing. Metabolites were collected in the supernatant by centrifugation at 14000 rpm for 5 min at 4°C. Previous step was repeated twice by adding 100µl 80% chilled solvent each time to increase metabolite yield and polled extracts were stored in -80°C refrigerator till further analysis.

Sampleprep ID:

SP001464

Sampleprep Summary:

Cell pellets were extracted with 200ul of chilled extraction solvent (80% MetOH in MS grade water containing 1ng/µl PIPES and U13C-U15N-glutamine as internal standard) was added to the pellet, mixed and incubated in ice for 5 minutes for quenching metabolism. This is followed by sonication in water bath for 15 min at 4°C with intermittent vortexing. Metabolites were collected in the supernatant by centrifugation at 14000 rpm for 5 min at 4°C. Previous step was repeated twice by adding 100µl 80% chilled solvent each time to increase metabolite yield and polled extracts were stored in -80°C refrigerator till further analysis.