A Protein Phosphatase 1 Specific Phatase Rgeting Eptide (PhosTAP) to Identify the PP1 Phosphatome

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2024 Oct 24

PMID 39446389

Authors

Meng S Choy

Hieu T Nguyen

Ganesan S Kumar

Wolfgang Peti

Arminja N Kettenbach

Rebecca Page

Affiliations

Soon will be listed here.

Abstract

Phosphoprotein phosphatases (PPPs) are the key serine/threonine phosphatases that regulate all essential signaling cascades. In particular, Protein Phosphatase 1 (PP1) dephosphorylates ~80% of all ser/thr phosphorylation sites. Here, we developed a phosphatase targeting peptide (PhosTAP) that binds all PP1 isoforms and does so with a stronger affinity than any other known PP1 regulator. This PhosTAP can be used as a PP1 recruitment tool for Phosphorylation Targeting Chimera (PhosTAC)-type recruitment in in vitro and cellular experiments, as well as in phosphoproteomics experiments to identify PP1-specific substrates and phosphosites. The latter is especially important to further our understanding of cellular signaling, as the identification of substrates and especially phosphosites that are targeted by specific phosphatases lags behind that of their kinase counterparts. Using PhosTAP-based proteomics, we show that, counter to our current understanding, many PP1 regulators are also substrates, that the number of residues between regulator PP1-binding and phosphosites vary significantly, and that PP1 counteracts the activities of mitotic kinases. Finally, we also found that Haspin kinase is a direct substrate of PP1 and that its PP1-dependent dephosphorylation modulates its activity during anaphase. Together, we show that PP1-specific PhosTAPs are a powerful tool for +studying PP1 activity in vitro and in cells.

Citing Articles

The phosphate of life.

De Vita E, Page R Nat Chem. 2025; 17(3):460.

PMID: 40050688 DOI: 10.1038/s41557-025-01758-3.

A protein phosphatase 1 specific phatase rgeting eptide (PhosTAP) to identify the PP1 phosphatome.

Choy M, Nguyen H, Kumar G, Peti W, Kettenbach A, Page R Proc Natl Acad Sci U S A. 2024; 121(44):e2415383121.

PMID: 39446389 PMC: 11536154. DOI: 10.1073/pnas.2415383121.

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