Simultaneously Revealing All Lysine Acylations in an Organism to Elucidate Substrate Metabolism
- Department of Biological Chemistry UCLA Los Angeles CA
Within the past 8 years, lysine acetylation knowledge has extended greatly beyond one modification important for chromatin maintenance and transcription regulation to the whole family of reversible conserved short‐chain acylations emerging as an important regulatory mechanism coordinating metabolism in response to changes in nutrient availability. Inspired by these lysine acylation discoveries, we devise a mass spectrometry (MS) method to simultaneously reveal and evaluate all lysine acylations in an organism. Hypothesizing that an organism's pool of acyl‐CoA esters could suggest potential acyl‐lysine modifications hidden within MS/MS datasets, we chose two syntrophic bacteria, Syntrophomonas wolfei and Syntrophus aciditrophicus, for the study. They activate fatty acid and/or aromatic substrates with coenzyme A and stepwise convert acyl‐CoAs down to acetyl‐CoA for ATP production, thus containing a variety of acyl‐CoA intermediates. Different from the current approach of immuno‐enriching acylated peptide prior to MS, our shotgun non‐enrichment MS approach allows a quick, non immuno‐bias, economical and quantitative one‐step assessment of all lysine acylations within an organism, and the modification stoichiometries and their relative levels. We also showed characteristic ions are reliable to detect and confirm lysine acylations, establishing seven lysine acylations first seen in any organism and doubling the number of known short‐chain acylations. Our lysine acylation studies suggest a metabolism regulation mechanism in these syntrophs living at the thermodynamic limit of feasibility. This proteomic approach can be applied to any organism having a complex profile of acyl‐CoAs, especially anaerobic bacteria degrading fatty acids and aromatic compounds and gut bacteria producing butyrate.
Support or Funding Information
This work is funded by the US Department of Energy (UCLA Institute of Genomics and Proteomics, DE‐FC03‐02ER63421 to J. Loo) and the National Institutes of Health (R01GM085402 to J. Loo and R. Ogorzalek Loo).
- Sponsoring Organization:
- USDOE
- Grant/Contract Number:
- FC03-02ER63421
- OSTI ID:
- 1787058
- Journal Information:
- FASEB Journal, Journal Name: FASEB Journal Journal Issue: S1 Vol. 30; ISSN 0892-6638
- Publisher:
- FASEBCopyright Statement
- Country of Publication:
- United States
- Language:
- English
Similar Records
Dynamic acylome reveals metabolite driven modifications in Syntrophomonas wolfei
Pyrophosphate-Dependent ATP Formation from Acetyl Coenzyme A in Syntrophus aciditrophicus, a New Twist on ATP Formation