PXD047986

PXD047986 is an original dataset announced via ProteomeXchange.

Title	Engineering cell-free systems by chemoproteomic-assisted phenotypic screening.
Description	Phenotypic screening is a valuable tool to both understand and engineer complex biological systems. We demonstrate the functionality of this approach in the development of cell-free protein synthesis (CFPS) technology. Phenotypic screening identified numerous compounds that enhanced protein production in yeast lysate CFPS reactions. Notably, many of these were competitive ATP kinase inhibitors, with exploitation of their inherent substrate promiscuity redirecting ATP flux towards heterologous protein expression. Chemoproteomic- guided strain engineering partially phenocopied drug effects, with a 30% increase in protein yield observed upon deletion of the ATP-consuming SSA1 component of the HSP70 chaperone. Moreover, drug-mediated metabolic rewiring coupled with template optimization generated the highest protein yields in yeast CFPS to date using a hitherto less efficient, but more cost-effective glucose energy regeneration system. Our approach highlights the utility of target-agnostic phenotypic screening and target identification to deconvolute cell-lysate complexity, adding to the expanding repertoire of strategies for improving CFPS.
HostingRepository	jPOST
AnnounceDate	2024-07-22
AnnouncementXML	Submission_2024-07-21_18:35:45.092.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Radoslaw Sobota
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	L-methionine sulfoxide; S-carboxamidomethyl-L-cysteine; alpha-amino acetylated residue; unknown modification; unknown modification; deamidated L-asparagine; deamidated L-glutamine
Instrument	instrument

Revision	Datetime	Status	ChangeLog Entry
0	2023-12-20 00:58:27	ID requested
⏵ 1	2024-07-21 18:35:45	announced

Levitskaya Z, Ser Z, Koh H, Mei WS, Chee S, Sobota RM, Ghadessy JF, Engineering cell-free systems by chemoproteomic-assisted phenotypic screening. RSC Chem Biol, 5(4):372-385(2024) [pubmed]

submitter keyword: Cell Free Protein synthesis, Proteomics, Thermal-assisted, Yeast

Radoslaw M. Sobota, John F. Ghadessy
lab head
Radoslaw Sobota
contact affiliation	IMCB A-STAR Singapore
dataset submitter

jPOST dataset URI

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