Rapid and Reversible Knockdown of Endogenously Tagged Endosomal Proteins via an Optimized HaloPROTAC Degrader.


Journal

ACS chemical biology
ISSN: 1554-8937
Titre abrégé: ACS Chem Biol
Pays: United States
ID NLM: 101282906

Informations de publication

Date de publication:
17 05 2019
Historique:
pubmed: 13 4 2019
medline: 7 1 2020
entrez: 13 4 2019
Statut: ppublish

Résumé

Inducing post-translational protein knockdown is an important approach to probe biology and validate drug targets. An efficient strategy to achieve this 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 DC

Identifiants

pubmed: 30978004
doi: 10.1021/acschembio.8b01016
pmc: PMC6528276
doi:

Substances chimiques

Proteins 0
Class III Phosphatidylinositol 3-Kinases EC 2.7.1.137
Protein Serine-Threonine Kinases EC 2.7.11.1

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

882-892

Subventions

Organisme : Medical Research Council
ID : MC_U127015387
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_UU_00018/1
Pays : United Kingdom
Organisme : Medical Research Council
ID : MC_UU_12016/2
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
Pays : United Kingdom

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Auteurs

Hannah Tovell (H)

Medical Research Council (MRC) Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences , University of Dundee , Dow Street , Dundee DD1 5EH , U.K.

Andrea Testa (A)

Division of Biological Chemistry and Drug Discovery, School of Life Sciences , University of Dundee , Dow Street , Dundee , DD1 5EH , U.K.

Chiara Maniaci (C)

Division of Biological Chemistry and Drug Discovery, School of Life Sciences , University of Dundee , Dow Street , Dundee , DD1 5EH , U.K.

Houjiang Zhou (H)

Medical Research Council (MRC) Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences , University of Dundee , Dow Street , Dundee DD1 5EH , U.K.

Alan R Prescott (AR)

Dundee Imaging Facility, School of Life Sciences , University of Dundee , Dundee DD1 5EH , U.K.

Thomas Macartney (T)

Medical Research Council (MRC) Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences , University of Dundee , Dow Street , Dundee DD1 5EH , U.K.

Alessio Ciulli (A)

Division of Biological Chemistry and Drug Discovery, School of Life Sciences , University of Dundee , Dow Street , Dundee , DD1 5EH , U.K.

Dario R Alessi (DR)

Medical Research Council (MRC) Protein Phosphorylation and Ubiquitylation Unit, School of Life Sciences , University of Dundee , Dow Street , Dundee DD1 5EH , U.K.

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Classifications MeSH