PXD012828

PXD012828 is an original dataset announced via ProteomeXchange.

Dataset Summary

Title	Rapid and reversible knockdown of endogenously tagged endosomal proteins via an optimized HaloPROTAC degrader
Description	Combining pharmacologic and genetic tools to induce post-translational protein knockdown is an emerging approach to probe biology and validate potential drug targets. A powerful strategy involves expression of a protein of interest fused to an exogenous tag, allowing tag-directed chemical degraders to mediate protein ubiquitylation and proteasomal degradation. Here, we combine improved HaloPROTAC degrader probes with CRISPR/Cas9 genome editing technology to trigger rapid degradation of endogenous target proteins. Our optimized probe, HaloPROTAC-E, a chloroalkane conjugate of high-affinity VHL binder VH298, induced reversible degradation of two endosomally-localized proteins, SGK3 and VPS34, with a DC50 of 3-10 nM. HaloPROTAC-E induced rapid (~50% degradation after 30 minutes) and complete (Dmax of ~95% at 48 h) depletion of Halo-tagged SGK3, blocking downstream phosphorylation of the SGK3 substrate NDRG1. This study exemplifies the combination of HaloPROTACs with CRISPR/Cas9 endogenous protein tagging as a useful method to induce rapid and reversible degradation of endogenous proteins to interrogate their function.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:53:26.080.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Houjiang Zhou
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue
Instrument	Orbitrap Fusion Lumos

Dataset History

Revision	Datetime	Status	ChangeLog Entry
0	2019-02-22 01:40:01	ID requested
1	2019-11-12 19:07:31	announced
⏵ 2	2024-10-22 04:53:33	announced	2024-10-22: Updated project metadata.

Publication List

Tovell H, Testa A, Maniaci C, Zhou H, Prescott AR, Macartney T, Ciulli A, Alessi DR, Rapid and Reversible Knockdown of Endogenously Tagged Endosomal Proteins via an Optimized HaloPROTAC Degrader. ACS Chem Biol, 14(5):882-892(2019) [pubmed]
10.1021/acschembio.8b01016;

Tovell H, Testa A, Maniaci C, Zhou H, Prescott AR, Macartney T, Ciulli A, Alessi DR, Rapid and Reversible Knockdown of Endogenously Tagged Endosomal Proteins via an Optimized HaloPROTAC Degrader. ACS Chem Biol, 14(5):882-892(2019) [pubmed]

10.1021/acschembio.8b01016;

Keyword List

curator keyword: Biomedical
submitter keyword: HaloPROTAC, VPS34, Degradation

curator keyword: Biomedical

submitter keyword: HaloPROTAC, VPS34, Degradation

Contact List

Dario Alessi
contact affiliation	1 Medical Research Council (MRC) Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences, University of Dundee
contact email	d.r.alessi@dundee.ac.uk
lab head
Houjiang Zhou
contact affiliation	University of Dundee
contact email	h.y.zhou@dundee.ac.uk
dataset submitter

Full Dataset Link List

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PRIDE project URI

Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2019/11/PXD012828

PRIDE project URI

Repository Record List

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