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.
01_22_18 HeLa P+12C 70kV CO2 12k neg 01.zip
03_05_18 HeLa P-12C Gly 70KV CO2 14k neg 01.zip
05_09_18 HeLa C13 P- 70kV CO2 13k neg 01.zip
05_09_18 HeLa N15 P- 70kV CO2 13k neg.zip
12_04_18 HeLa knockout 15N 70kv CO214k neg.zip
12_20_17 HeLa P+13C 70kv CO2 12k neg.zip
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