Dissecting the sequence determinants for dephosphorylation by the catalytic subunits of phosphatases PP1 and PP2A.

Nature Communications
Bernhard HoermannMaja Köhn

Abstract

The phosphatases PP1 and PP2A are responsible for the majority of dephosphorylation reactions on phosphoserine (pSer) and phosphothreonine (pThr), and are involved in virtually all cellular processes and numerous diseases. The catalytic subunits exist in cells in form of holoenzymes, which impart substrate specificity. The contribution of the catalytic subunits to the recognition of substrates is unclear. By developing a phosphopeptide library approach and a phosphoproteomic assay, we demonstrate that the specificity of PP1 and PP2A holoenzymes towards pThr and of PP1 for basic motifs adjacent to the phosphorylation site are due to intrinsic properties of the catalytic subunits. Thus, we dissect this amino acid specificity of the catalytic subunits from the contribution of regulatory proteins. Furthermore, our approach enables discovering a role for PP1 as regulator of the GRB-associated-binding protein 2 (GAB2)/14-3-3 complex. Beyond this, we expect that this approach is broadly applicable to detect enzyme-substrate recognition preferences.

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Citations

Jun 9, 2021·Biochemical Society Transactions·Bernhard Hoermann, Maja Köhn

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Datasets Mentioned

BETA
GENA934

Methods Mentioned

BETA
size-exclusion chromatography
gel-filtration
co-IP
phage display
PCR
transfections
immunoprecipitation
transfection
Assay
acetylation

Software Mentioned

IsobarQuant
Process
Gen5
R
PhosphoSitePlus
EmpCorr
Fiji
MaxQuant
uniprot
Needle

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