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.
|Work Title||Metabolites imaging in single HeLa cells using GCIB-SIMS|
|License||Public Domain Mark 1.0|
|Deposited||February 26, 2020|
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