PXD012964

PXD012964 is an original dataset announced via ProteomeXchange.

Title	Measuring glycolytic flux in single yeast cells with an orthogonal synthetic biosensor
Description	Metabolic heterogeneity modulates productivity, antibiotic resistance and cancer aggressiveness. Since metabolic fluxes represent the functional output of metabolism, with glycolytic flux correlating with highly-productive phenotypes and cancer, such flux map will be indicative of the cellular metabolic state. Therefore, the quantification of metabolic fluxes is vital to identify the existence of metabolic subpopulations and to understand the process of their emergence at the single-cell level. However, so far inference of metabolic fluxes in individual cells is not possible as no method is available. Here, we developed a biosensor for glycolytic flux measurements in single yeast cells drawing on the robust correlation between fructose-1,6-bisphosphate (FBP) and flux levels in yeast, and using the B. subtilis FBP-binding transcription factor CggR. We followed a systematic engineering approach starting from promoter design, computational protein design and protein engineering, accompanied by strict characterization of the biosensor using different biochemical methods, proteomics, metabolomics and physiological analyses. As proof of principle, we applied the biosensor in vivo in the search for metabolic subpopulations in yeast cultures and, using fluorescence microscopy, we demonstrated that quiescent yeast cells have low glycolytic fluxes in comparison to coexisting dividing cells. We anticipate that our biosensor will contribute with unprecedented resolution for the study of metabolic subpopulations, to understand how and why metabolic subpopulations emerge and, very importantly, give clues on how to counteract the undesirable effects of such.
HostingRepository	PRIDE
AnnounceDate	2019-12-17
AnnouncementXML	Submission_2020-01-15_05:41:15.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD012964
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Alexander Schmidt
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	Oxidation; Acetyl; Carbamidomethyl
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2019-03-05 03:22:32	ID requested
1	2019-12-17 08:26:22	announced
⏵ 2	2020-01-15 05:41:17	announced	2020-01-15: Updated publication reference for PubMed record(s): 31885198.

Monteiro F, Hubmann G, Takhaveev V, Vedelaar SR, Norder J, Hekelaar J, Saldida J, Litsios A, Wijma HJ, Schmidt A, Heinemann M, Measuring glycolytic flux in single yeast cells with an orthogonal synthetic biosensor. Mol Syst Biol, 15(12):e9071(2019) [pubmed]

curator keyword: Biological

submitter keyword: Yeast, mCherry, CggR

Alexander Schmidt
contact affiliation	Biozentrum, University of Basel, 4056 Basel, Switzerland
contact email	alex.schmidt@unibas.ch
lab head
Alexander Schmidt
contact affiliation	Proteomics Core Facility
contact email	alex.schmidt@unibas.ch
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD012964
     1. Label: PRIDE project
     2. Name: Measuring glycolytic flux in single yeast cells with an orthogonal synthetic biosensor