Metabolites imaging in single HeLa cells using GCIB-SIMS

Metabolons, multiprotein complexes consisting of sequential enzymes of a metabolic pathway, are proposed to be biosynthetic ‘hot-spots’ within the cell. However, experimental demonstration of their presence and features of their function has remained challenging. We used metabolomics and in-situ three-dimensional sub-micron chemical imaging of single cells by gas cluster ion beam secondary ion mass spectrometry (GCIB-SIMS) to directly visualize de novo purine biosynthesis (DNPB) by a multienzyme complex, the purinosome. We found that purinosomes comprise nine enzymes that act synergistically, channeling the pathway intermediates to synthesize purine nucleotides, increasing the pathway flux, and influencing the AMP:GMP ratio. Our work also highlights application of high-resolution GCIB-SIMS for multiplexed biomolecular analysis at the level of single cells.

Files

Metadata

Work Title Metabolites imaging in single HeLa cells using GCIB-SIMS
Access
Open Access
Creators
  1. Vidhi Pareek
  2. Hua Tian
  3. Nicholas Winograd
  4. Stephen James Benkovic
Keyword
  1. de novo purine biosynthesis
  2. GCIB-SIMS
  3. 3D chemical imaging
  4. Purinosome
  5. metabolon
License Public Domain Mark 1.0
Work Type Dataset
Publication Date 2020
DOI doi:10.26207/8rm3-g809
Deposited February 26, 2020

Versions

Analytics

Collections

This resource is currently not in any collection.

Work History

Version 1
published

  • Created
  • Added 12_20_17_HeLa_P_13C_70kv_CO2_12k_neg.zip
  • Added README.txt
  • Added 05_09_18_HeLa_C13_P-_70kV_CO2_13k_neg_01.zip
  • Added 03_05_18_HeLa_P-12C_Gly_70KV_CO2_14k_neg_01.zip
  • Added 05_09_18_HeLa_N15_P-_70kV_CO2_13k_neg.zip
  • Added 01_22_18_HeLa_P_12C_70kV_CO2_12k_neg_01.zip
  • Added 12_04_18_HeLa_knockout_15N_70kv_CO214k_neg.zip
  • Added Creator Vidhi Pareek
  • Added Creator Hua Tian
  • Added Creator Nicholas Winograd
  • Added Creator Stephen James Benkovic
  • Published