Dataset Information

An affinity-directed phosphatase (AdPhosphatase) system for targeted protein dephosphorylation

ABSTRACT: Reversible protein phosphorylation, catalysed by protein kinases and phosphatases, is a fundamental process that controls protein function and intracellular signalling. Failure of phospho-control accounts for many human diseases. While a kinase phosphorylates multiple substrates, a substrate is often phosphorylated by multiple kinases. This renders phospho control at the substrate level challenging, as it requires inhibition of multiple kinases, which would thus affect other kinase substrates. Here, we describe the development and application of the affinity-directed phosphatase (AdPhosphatase) system for targeted dephosphorylation of specific phospho-substrates. By deploying the Protein Phosphatase 1 or 2A catalytic subunits conjugated to an antigen-stabilised anti-GFP nanobody, we can promote the dephosphorylation of two independent phospho-proteins, FAM83D or ULK1, knocked in with GFP-tags using CRISPR/Cas9, with exquisite specificity. By redirecting protein phosphatases to neo-substrates through nanobody-mediated proximity, AdPhosphatase can alter the phospho-status and function of target proteins and thus, offers a new modality for potential drug discovery approaches.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Epithelial Cell

SUBMITTER: Gopal Sapkota

LAB HEAD: Professor Gopal Sapkota

PROVIDER: PXD039200 | Pride | 2023-02-01

REPOSITORIES: Pride

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An affinity-directed phosphatase, AdPhosphatase, system for targeted protein dephosphorylation.

Simpson Luke M LM Fulcher Luke J LJ Sathe Gajanan G Brewer Abigail A Zhao Jin-Feng JF Squair Daniel R DR Crooks Jennifer J Wightman Melanie M Wood Nicola T NT Gourlay Robert R Varghese Joby J Soares Renata F RF Sapkota Gopal P GP

Cell chemical biology 20230130 2

Reversible protein phosphorylation, catalyzed by protein kinases and phosphatases, is a fundamental process that controls protein function and intracellular signaling. Failure of phospho-control accounts for many human diseases. While a kinase phosphorylates multiple substrates, a substrate is often phosphorylated by multiple kinases. This renders phospho-control at the substrate level challenging, as it requires inhibition of multiple kinases, which would thus affect other kinase substrates. He ...[more]

PMID: 36720221

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			Action	DRS
	FAM10_PHOSPHO.raw	Raw
	FAM11_PHOSPHO.raw	Raw
	FAM12_PHOSPHO.raw	Raw
	FAM1_PHOSPHO.raw	Raw
	FAM2_PHOSPHO.raw	Raw