How does phosphorylation affect interaction between 14-3-3ζ and
Tau proteins?

a 2024

How does phosphorylation affect interaction between 14-3-3ζ and Tau proteins?

KOZELEKOVÁ, Aneta, Lucia IĽKOVIČOVÁ, Radek CRHA, Alena HOFROVÁ, Jozef HRITZ et. al.

Základní údaje

Originální název

How does phosphorylation affect interaction between 14-3-3ζ and Tau proteins?

Název česky

Jak ovlivňuje fosforylace interakci mezi proteiny 14-3-3ζ a Tau?

Autoři

KOZELEKOVÁ, Aneta, Lucia IĽKOVIČOVÁ, Radek CRHA, Alena HOFROVÁ a Jozef HRITZ

Vydání

XX Discussions in Structural Molecular Biology and 7th User Meeting of CIISB, 2024

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Organizační jednotka

Středoevropský technologický institut

ISSN

Příznaky

Mezinárodní význam

Změněno: 12. 7. 2024 09:36, Mgr. Aneta Kozeleková

Anotace

V originále

Phosphorylation is a post-translational modification that affects structure, function, and interactions of proteins. 14-3-3ζ protein, an abundant human regulatory protein, in non-phosphorylated state exists as a dimer [1]. However, after phosphorylation at Ser58 (pS58), it monomerizes and changes its properties [2, 3]. Hyperphosphorylation of Tau protein, a microtubule-associated protein, causes detachment of Tau from microtubules in neurons and leads to neurodegeneration [4]. Hyperphosphorylated Tau aggregates into neurofibrillary tangles (NFTs) - one of the hallmarks of Alzheimer’s disease (AD). As 14-3-3ζ proteins were found colocalized in the NFTs [5], their interconnection with Tau in AD needs to be comprehended. In our study, we aimed to compare interaction properties of dimeric 14-3-3ζ WT and monomeric 14-3-3ζ pS58 with respect to Tau protein. The interaction with Tau protein phosphorylated by protein kinase A (PKA) was inspected from various points of view. The binding affinity, stoichiometry, and interacting residues were studied using native-PAGE, chemical cross-linking, tandem MS, and NMR spectroscopy. We revealed that phosphorylation of 14-3-3ζ at Ser58 decreases its affinity to Tau protein and changes binding stoichiometry. Both NMR and cross-linking results suggested that Tau is in contact with 14-3-3ζ proteins via the proline-rich domain and microtubule-binding domain. Moreover, cross-linking data showed that not only the binding channel of 14-3-3ζ protein is responsible for Tau binding, but also the outer 14-3-3ζ protein surface and exposed dimeric interface of monomeric 14-3-3ζ pS58 are involved. In summary, we provide novel insight into the 14-3-3ζ+Tau interaction and its regulation by phosphorylation of both partners. 1. V. Obsilova & T. Obsil, Front. Mol. Biosci., 9, (2022), 1-15. 2. A. Kozeleková, A. Náplavová, T. Brom, N. Gašparik, J. Šimek, J. Houser, J. Hritz, Front. Chem., 10, (2022), 1-17. 3. Z. Trošanová, P. Louša, A. Kozeleková, T. Brom, N. Gašparik, J. Tungli, V. Weisová, E. Župa, G. Žoldák, J. Hritz, J. Mol. Biol., 434, (2022), 167479. 4. T. Arendt, J. T. Stieler, M. Holzer, Brain. Res. Bull., 126, (2016), 238-292. 5. R. Layfield, J. Fergusson, A. Aitken, J. Lowe, M. Landon, R. J. Mayer, Neurosci. Lett., 209, (1996), 57-60. This project has received funding from the European Union’s Horizon Europe program under the grant agreement No. 101087124 and from Czech Science Foundation (GF20-05789L). We acknowledge CEITEC Proteomics Core Facility and Josef Dadok National NMR Centre of CIISB, Instruct-CZ Centre, supported by MEYS CR (LM2023042) and European Regional Development Fund-Project „UP CIISB“ (No. CZ.02.1.01/0.0/0.0/18_046/0015974). Computational resources were provided by the e-INFRA CZ project (ID:90254), supported by MEYS CR.

Návaznosti

EF18_046/0015974, projekt VaV

Název: Modernizace České infrastruktury pro integrativní strukturní biologii

GF20-05789L, projekt VaV

Název: Charakterizace přirozeně neuspořádaných proteinů

Investor: Grantová agentura ČR, Characterization of intrinsically disordered proteins, Partnerská agentura (Rakousko)

LM2023042, projekt VaV

Název: Česká infrastruktura pro integrativní strukturní biologii

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CIISB - Česká infrastruktura pro integrativní strukturní biologii

101087124, interní kód MU

Název: Alzheimer's Disease Diagnostics Innovation and Translation to Clinical Practice in Central Europe

Investor: Evropská unie, Alzheimer's Disease Diagnostics Innovation and Translation to Clinical Practice in Central Europe, Rozšiřování účasti a posílení ERA

90254, velká výzkumná infrastruktura

Název: e-INFRA CZ II

Citovat

KOZELEKOVÁ, Aneta, Lucia IĽKOVIČOVÁ, Radek CRHA, Alena HOFROVÁ a Jozef HRITZ. How does phosphorylation affect interaction between 14-3-3ζ and Tau proteins? In XX Discussions in Structural Molecular Biology and 7th User Meeting of CIISB. 2024. ISSN 1211-5894.

@proceedings{2418110,
   author = {Kozeleková, Aneta and Iľkovičová, Lucia and Crha, Radek and Hofrová, Alena and Hritz, Jozef},
   booktitle = {XX Discussions in Structural Molecular Biology and 7th User Meeting of CIISB},
   language = {eng},
   title = {How does phosphorylation affect interaction between 14-3-3ζ and Tau proteins?},
   url = {https://cssb.structbio.org/xx-discussions-in-structural-molecular-biology-and-7th-user-meeting-of-ciisb-czech-infrastructure-for-integrative-structural-biologyp/},
   year = {2024}
}

TY  - CONF
ID  - 2418110
AU  - Kozeleková, Aneta - Iľkovičová, Lucia - Crha, Radek - Hofrová, Alena - Hritz, Jozef
PY  - 2024
TI  - How does phosphorylation affect interaction between 14-3-3ζ and Tau proteins?
UR  - https://cssb.structbio.org/xx-discussions-in-structural-molecular-biology-and-7th-user-meeting-of-ciisb-czech-infrastructure-for-integrative-structural-biologyp/
N2  - Phosphorylation is a post-translational modification that affects structure, function, and interactions of proteins. 14-3-3ζ protein, an abundant human regulatory protein, in non-phosphorylated state exists as a dimer [1]. However, after phosphorylation at Ser58 (pS58), it monomerizes and changes its properties [2, 3]. Hyperphosphorylation of Tau protein, a microtubule-associated protein, causes detachment of Tau from microtubules in neurons and leads to neurodegeneration [4]. Hyperphosphorylated Tau aggregates into neurofibrillary tangles (NFTs) - one of the hallmarks of Alzheimer’s disease (AD). As 14-3-3ζ proteins were found colocalized in the NFTs [5], their interconnection with Tau in AD needs to be comprehended. In our study, we aimed to compare interaction properties of dimeric 14-3-3ζ WT and monomeric 14-3-3ζ pS58 with respect to Tau protein. The interaction with Tau protein phosphorylated by protein kinase A (PKA) was inspected from various points of view. The binding affinity, stoichiometry, and interacting residues were studied using native-PAGE, chemical cross-linking, tandem MS, and NMR spectroscopy. We revealed that phosphorylation of 14-3-3ζ at Ser58 decreases its affinity to Tau protein and changes binding stoichiometry. Both NMR and cross-linking results suggested that Tau is in contact with 14-3-3ζ proteins via the proline-rich domain and microtubule-binding domain. Moreover, cross-linking data showed that not only the binding channel of 14-3-3ζ protein is responsible for Tau binding, but also the outer 14-3-3ζ protein surface and exposed dimeric interface of monomeric 14-3-3ζ pS58 are involved. In summary, we provide novel insight into the 14-3-3ζ+Tau interaction and its regulation by phosphorylation of both partners. 1. V. Obsilova & T. Obsil, Front. Mol. Biosci., 9, (2022), 1-15. 2. A. Kozeleková, A. Náplavová, T. Brom, N. Gašparik, J. Šimek, J. Houser, J. Hritz, Front. Chem., 10, (2022), 1-17. 3. Z. Trošanová, P. Louša, A. Kozeleková, T. Brom, N. Gašparik, J. Tungli, V. Weisová, E. Župa, G. Žoldák, J. Hritz, J. Mol. Biol., 434, (2022), 167479. 4. T. Arendt, J. T. Stieler, M. Holzer, Brain. Res. Bull., 126, (2016), 238-292. 5. R. Layfield, J. Fergusson, A. Aitken, J. Lowe, M. Landon, R. J. Mayer, Neurosci. Lett., 209, (1996), 57-60. This project has received funding from the European Union’s Horizon Europe program under the grant agreement No. 101087124 and from Czech Science Foundation (GF20-05789L). We acknowledge CEITEC Proteomics Core Facility and Josef Dadok National NMR Centre of CIISB, Instruct-CZ Centre, supported by MEYS CR (LM2023042) and European Regional Development Fund-Project „UP CIISB“ (No. CZ.02.1.01/0.0/0.0/18_046/0015974). Computational resources were provided by the e-INFRA CZ project (ID:90254), supported by MEYS CR.
ER  -

KOZELEKOVÁ, Aneta, Lucia IĽKOVIČOVÁ, Radek CRHA, Alena HOFROVÁ a Jozef HRITZ. How does phosphorylation affect interaction between 14-3-3ζ and Tau proteins? In \textit{XX Discussions in Structural Molecular Biology and 7th User Meeting of CIISB}. 2024. ISSN~1211-5894.

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