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PXD045052-1

PXD045052 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleRegulation of eDHFR-tagged proteins with trimethoprim PROTACs
DescriptionTemporal control of proteins in cells and living animals is crucial to improving the understanding of protein function in the post-genomic era. In addition, technologies that offer such control for engineered proteins could be used in therapeutic applications. Since regulation of proteins at a genomic or transcriptional level can be irreversible or a slow process, these tools may not be useful in settings where rapid temporal control is required to achieve immediate knockdown effects. PRoteolysis-TArgeting Chimeras (PROTACs) have emerged as a strategy to achieve rapid, post-translational control of protein abundance via recruitment of an E3 ligase to the target protein of interest. Here, we developed several PROTAC molecules by covalently linking the antibiotic trimethoprim (TMP) to pomalidomide, a small molecule ligand of the E3 ligase Cereblon. These molecules induce degradation of various proteins of interest (POIs) genetically fused to E. coli dihydrofolate reductase (eDHFR), the molecular target of TMP. We demonstrate that various eDHFR-tagged proteins, from fluorescent proteins to transcription factors and membrane-associated proteins, can be downregulated to 95% of maximum expression with our lead PROTAC molecule 7c. The data suggest that TMP-based PROTACs induce maximal degradation of POIs at drug concentrations that minimally affect the expression of immunomodulatory imide drug (IMiD)-sensitive neosubstrates. Finally, we show the ability to achieve multiplexed regulation with another known degron-PROTAC pair, and the formidable strength of our system for reversible protein regulation in a rodent model of metastatic cancer. Altogether, TMP PROTACs are a robust approach for selective and reversible degradation of eDHFR-tagged protein and have a strong potential for translation to in vivo models as well as dual degradation strategies with existing technologies.
HostingRepositoryPRIDE
AnnounceDate2023-12-06
AnnouncementXMLSubmission_2023-12-06_14:31:59.971.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterRichard Lauman
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListNo PTMs are included in the dataset
InstrumentQ Exactive HF
Dataset History
RevisionDatetimeStatusChangeLog Entry
02023-09-02 20:26:57ID requested
12023-12-06 14:32:00announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: eDHFR, Degradation, PROTACs
Contact List
Richard Lauman
contact affiliationDepartment of Radiology Perelman School of Medicine at the University of Pennsylvania
contact emailmark.sellmyer@pennmedicine.upenn.edu
lab head
Richard Lauman
contact affiliationUniverstiy of Pennsylvania
contact emailrichard.lauman@gmail.com
dataset submitter
Full Dataset Link List
Dataset FTP location
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