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.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
| 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 |
| 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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Combined analysis:
| Analysis ID | AN002303 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Accela 1250 |
| Column | Thermo Accucore C18 (100 x 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 x 2.1mm,2.6um) |
| Flow Gradient: | 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. |
| Flow Rate: | 200 µl/min |
| Solvent A: | 100% water; 0.1 % formic acid |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | Reversed phase |
