Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification

Research output: Contribution to journal › Journal article › Research › peer-review

Standard

Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification. / Hein, Jamin B.; Nguyen, Hieu T.; Garvanska, Dimitriya H.; Nasa, Isha; Kruse, Thomas; Feng, Yinnian; Lopez Mendez, Blanca; Davey, Norman; Kettenbach, Arminja N.; Fordyce, Polly M.; Nilsson, Jakob.

In: Molecular Systems Biology, Vol. 19, e11782, 2023.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Hein, JB, Nguyen, HT, Garvanska, DH, Nasa, I, Kruse, T, Feng, Y, Lopez Mendez, B, Davey, N, Kettenbach, AN, Fordyce, PM & Nilsson, J 2023, 'Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification', Molecular Systems Biology, vol. 19, e11782. https://doi.org/10.15252/msb.202311782

APA

Hein, J. B., Nguyen, H. T., Garvanska, D. H., Nasa, I., Kruse, T., Feng, Y., Lopez Mendez, B., Davey, N., Kettenbach, A. N., Fordyce, P. M., & Nilsson, J. (2023). Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification. Molecular Systems Biology, 19, [e11782]. https://doi.org/10.15252/msb.202311782

Vancouver

Hein JB, Nguyen HT, Garvanska DH, Nasa I, Kruse T, Feng Y et al. Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification. Molecular Systems Biology. 2023;19. e11782. https://doi.org/10.15252/msb.202311782

Author

Hein, Jamin B. ; Nguyen, Hieu T. ; Garvanska, Dimitriya H. ; Nasa, Isha ; Kruse, Thomas ; Feng, Yinnian ; Lopez Mendez, Blanca ; Davey, Norman ; Kettenbach, Arminja N. ; Fordyce, Polly M. ; Nilsson, Jakob. / Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification. In: Molecular Systems Biology. 2023 ; Vol. 19.

Bibtex

@article{2d10d3cae9cd48309e76179b12055fda,
title = "Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification",
abstract = "Phosphoprotein phosphatases (PPPs) regulate major signaling pathways, but the determinants of phosphatase specificity are poorly understood. This is because methods to investigate this at scale are lacking. Here, we develop a novel in vitro assay, MRBLE:Dephos, that allows multiplexing of dephosphorylation reactions to determine phosphatase preferences. Using MRBLE:Dephos, we establish amino acid preferences of the residues surrounding the dephosphorylation site for PP1 and PP2A-B55, which reveals common and unique preferences. To compare the MRBLE:Dephos results to cellular substrates, we focused on mitotic exit that requires extensive dephosphorylation by PP1 and PP2A-B55. We use specific inhibition of PP1 and PP2A-B55 in mitotic exit lysates coupled with phosphoproteomics to identify more than 2,000 regulated sites. Importantly, the sites dephosphorylated during mitotic exit reveal key signatures that are consistent with MRBLE:Dephos. Furthermore, integration of our phosphoproteomic data with mitotic interactomes of PP1 and PP2A-B55 provides insight into how binding of phosphatases to substrates shapes dephosphorylation. Collectively, we develop novel approaches to investigate protein phosphatases that provide insight into mitotic exit regulation.",
keywords = "mitotic exit, MRBLE-Pep, MRBLE:Dephos, protein phosphatase, substrates",
author = "Hein, {Jamin B.} and Nguyen, {Hieu T.} and Garvanska, {Dimitriya H.} and Isha Nasa and Thomas Kruse and Yinnian Feng and {Lopez Mendez}, Blanca and Norman Davey and Kettenbach, {Arminja N.} and Fordyce, {Polly M.} and Jakob Nilsson",
note = "Publisher Copyright: {\textcopyright} 2023 The Authors. Published under the terms of the CC BY 4.0 license.",
year = "2023",
doi = "10.15252/msb.202311782",
language = "English",
volume = "19",
journal = "Molecular Systems Biology",
issn = "1744-4292",
publisher = "Wiley-Blackwell",

}

RIS

TY - JOUR

T1 - Phosphatase specificity principles uncovered by MRBLE:Dephos and global substrate identification

AU - Hein, Jamin B.

AU - Nguyen, Hieu T.

AU - Garvanska, Dimitriya H.

AU - Nasa, Isha

AU - Kruse, Thomas

AU - Feng, Yinnian

AU - Lopez Mendez, Blanca

AU - Davey, Norman

AU - Kettenbach, Arminja N.

AU - Fordyce, Polly M.

AU - Nilsson, Jakob

N1 - Publisher Copyright: © 2023 The Authors. Published under the terms of the CC BY 4.0 license.

PY - 2023

Y1 - 2023

N2 - Phosphoprotein phosphatases (PPPs) regulate major signaling pathways, but the determinants of phosphatase specificity are poorly understood. This is because methods to investigate this at scale are lacking. Here, we develop a novel in vitro assay, MRBLE:Dephos, that allows multiplexing of dephosphorylation reactions to determine phosphatase preferences. Using MRBLE:Dephos, we establish amino acid preferences of the residues surrounding the dephosphorylation site for PP1 and PP2A-B55, which reveals common and unique preferences. To compare the MRBLE:Dephos results to cellular substrates, we focused on mitotic exit that requires extensive dephosphorylation by PP1 and PP2A-B55. We use specific inhibition of PP1 and PP2A-B55 in mitotic exit lysates coupled with phosphoproteomics to identify more than 2,000 regulated sites. Importantly, the sites dephosphorylated during mitotic exit reveal key signatures that are consistent with MRBLE:Dephos. Furthermore, integration of our phosphoproteomic data with mitotic interactomes of PP1 and PP2A-B55 provides insight into how binding of phosphatases to substrates shapes dephosphorylation. Collectively, we develop novel approaches to investigate protein phosphatases that provide insight into mitotic exit regulation.

AB - Phosphoprotein phosphatases (PPPs) regulate major signaling pathways, but the determinants of phosphatase specificity are poorly understood. This is because methods to investigate this at scale are lacking. Here, we develop a novel in vitro assay, MRBLE:Dephos, that allows multiplexing of dephosphorylation reactions to determine phosphatase preferences. Using MRBLE:Dephos, we establish amino acid preferences of the residues surrounding the dephosphorylation site for PP1 and PP2A-B55, which reveals common and unique preferences. To compare the MRBLE:Dephos results to cellular substrates, we focused on mitotic exit that requires extensive dephosphorylation by PP1 and PP2A-B55. We use specific inhibition of PP1 and PP2A-B55 in mitotic exit lysates coupled with phosphoproteomics to identify more than 2,000 regulated sites. Importantly, the sites dephosphorylated during mitotic exit reveal key signatures that are consistent with MRBLE:Dephos. Furthermore, integration of our phosphoproteomic data with mitotic interactomes of PP1 and PP2A-B55 provides insight into how binding of phosphatases to substrates shapes dephosphorylation. Collectively, we develop novel approaches to investigate protein phosphatases that provide insight into mitotic exit regulation.

KW - mitotic exit

KW - MRBLE-Pep

KW - MRBLE:Dephos

KW - protein phosphatase

KW - substrates

U2 - 10.15252/msb.202311782

DO - 10.15252/msb.202311782

M3 - Journal article

C2 - 37916966

AN - SCOPUS:85175715683

VL - 19

JO - Molecular Systems Biology

JF - Molecular Systems Biology

SN - 1744-4292

M1 - e11782

ER -

ID: 372966516