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 IDST001382
Study TitleDistinct metabolic states of a cell guide alternate fates of mutational buffering through altered proteostasis
Study SummaryChanges 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 NameShanmugam
First NameDhanasekaran
AddressDr. Homi Bhabha Road, Pune, maharashtra, 411008, India
Emaild.shanmugam@ncl.res.in
Phone2025902719
Submit Date2020-05-14
Raw Data AvailableYes
Raw Data File Type(s)mzXML
Analysis Type DetailLC-MS
Release Date2020-06-01
Release Version1
Dhanasekaran Shanmugam Dhanasekaran Shanmugam
https://dx.doi.org/10.21228/M8DM63
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Combined analysis:

Analysis ID AN002303
Analysis type MS
Chromatography type Reversed phase
Chromatography system Thermo Accela 1250
Column Thermo Accucore C18 (100 2.1mm, 2.6um)
MS Type ESI
MS instrument type Orbitrap
MS instrument name Thermo Q Exactive Orbitrap
Ion Mode NEGATIVE
Units Peak Intensity

Chromatography:

Chromatography ID:CH001692
Chromatography Summary:The acetonitrile:water extracts from parasites were dried under nitrogen flow and resuspended in 200 µls of water:methanol (97:3) containing 10 mM tributylamine and 15 mM acetic acid. A Thermo Accucore C18 column with a bed volume of 150 mm x 2.1 mm and 2.6 µ particle size was used. A solvent system composed of water buffered with 0.1 % formic acid (buffer A) and acetonitrile (buffer B), was used on a 20 minute gradient run with a flow rate of 200 µl/min as follows- hold at 10% acetonitrile for 30 seconds and gradually ramp up to 15%, 20%, 50%, 60% and 90% acetonitrile by 3, 6, 10, 12, 13 minutes, hold at 90% acetonitrile till 15 minutes, ramp down to 10% acetonitrile by 15.5 minutes and hold till 20 minutes.
Instrument Name:Thermo Accela 1250
Column Name:Thermo Accucore C18 (100 2.1mm, 2.6um)
Chromatography Type:Reversed phase
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