Preparation of Alzheimer's-disease-relevant Tau protein in
HEK293 mammalian cells

a 2024

Preparation of Alzheimer's-disease-relevant Tau protein in HEK293 mammalian cells

KOZELEKOVÁ, Aneta, Marie CHOVANČÍKOVÁ, Petr KAŠPÁREK, Helena REUTOVÁ, Martina HLOŽÁNKOVÁ et. al.

Základní údaje

Originální název

Preparation of Alzheimer's-disease-relevant Tau protein in HEK293 mammalian cells

Název česky

Příprava proteinu Tau relevantního pro Alzheimerovu chorobu v savčích HEK293 buňkách

Autoři

KOZELEKOVÁ, Aneta, Marie CHOVANČÍKOVÁ, Petr KAŠPÁREK, Helena REUTOVÁ, Martina HLOŽÁNKOVÁ a Jozef HRITZ

Vydání

CEITEC PhD School Conference 2024, Brno, 2024

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Organizační jednotka

Středoevropský technologický institut

Změněno: 12. 7. 2024 09:48, Mgr. Aneta Kozeleková

Anotace

V originále

Tau protein is a microtubule-binding protein that is usually post-translationally modified (PTM). Hyperphosphorylation and abnormal PTMs of Tau have been identified as key determinants connecting Tau and Alzheimer’s disease. After such abnormal modification, Tau detaches from microtubules and aggregates into neurotoxic neurofibrillary tangles. However, the molecular mechanism and exact factors causing Tau aggregation are so far not well understood. Here, we aimed to prepare Tau protein in mammalian HEK293 cells to obtain Tau bearing natural PTMs. Tau protein was successfully expressed, purified from the cell lysate, and its purity and identity were verified using SDS-PAGE, Western blot, and mass spectrometry (MS). Currently, the presence of PTMs within Tau protein is inspected by advanced MS analysis. Next step will be to study the effects of the PTMs on Tau structural, interaction, and aggregation properties in comparison to un-modified Tau expressed in E. coli or in vitro phosphorylated Tau (already published in article by Crha R, Kozeleková A, Hofrová A, Iľkovičová L, Gašparik N, Kadeřávek P, Hritz J. Hiding in plain sight: Complex interaction patterns between Tau and 14-3-3ζ protein variants. Int J Biol Macromol. 2024, doi: 10.1016/j.ijbiomac.2024.130802). The majority of results were obtained within Industrial PhD programme in biotech company BioVendor. Project acknowledgement: This project has received funding from the European Union’s Horizon Europe program under the grant agreement No. 101087124. We acknowledge CEITEC Proteomics Core Facility of CIISB, Instruct-CZ Centre, supported by MEYS CR (LM2023042, e-INFRA CZ (ID:90254)).

Návaznosti

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, Marie CHOVANČÍKOVÁ, Petr KAŠPÁREK, Helena REUTOVÁ, Martina HLOŽÁNKOVÁ a Jozef HRITZ. Preparation of Alzheimer's-disease-relevant Tau protein in HEK293 mammalian cells. In CEITEC PhD School Conference 2024, Brno. 2024.

@proceedings{2418111,
   author = {Kozeleková, Aneta and Chovančíková, Marie and Kašpárek, Petr and Reutová, Helena and Hložánková, Martina and Hritz, Jozef},
   booktitle = {CEITEC PhD School Conference 2024, Brno},
   language = {eng},
   title = {Preparation of Alzheimer's-disease-relevant Tau protein in HEK293 mammalian cells},
   year = {2024}
}

TY  - CONF
ID  - 2418111
AU  - Kozeleková, Aneta - Chovančíková, Marie - Kašpárek, Petr - Reutová, Helena - Hložánková, Martina - Hritz, Jozef
PY  - 2024
TI  - Preparation of Alzheimer's-disease-relevant Tau protein in HEK293 mammalian cells
N2  - Tau protein is a microtubule-binding protein that is usually post-translationally modified (PTM). Hyperphosphorylation and abnormal PTMs of Tau have been identified as key determinants connecting Tau and Alzheimer’s disease. After such abnormal modification, Tau detaches from microtubules and aggregates into neurotoxic neurofibrillary tangles. However, the molecular mechanism and exact factors causing Tau aggregation are so far not well understood. Here, we aimed to prepare Tau protein in mammalian HEK293 cells to obtain Tau bearing natural PTMs. Tau protein was successfully expressed, purified from the cell lysate, and its purity and identity were verified using SDS-PAGE, Western blot, and mass spectrometry (MS). Currently, the presence of PTMs within Tau protein is inspected by advanced MS analysis. Next step will be to study the effects of the PTMs on Tau structural, interaction, and aggregation properties in comparison to un-modified Tau expressed in E. coli or in vitro phosphorylated Tau (already published in article by Crha R, Kozeleková A, Hofrová A, Iľkovičová L, Gašparik N, Kadeřávek P, Hritz J. Hiding in plain sight: Complex interaction patterns between Tau and 14-3-3ζ protein variants. Int J Biol Macromol. 2024, doi: 10.1016/j.ijbiomac.2024.130802). The majority of results were obtained within Industrial PhD programme in biotech company BioVendor. Project acknowledgement: This project has received funding from the European Union’s Horizon Europe program under the grant agreement No. 101087124. We acknowledge CEITEC Proteomics Core Facility of CIISB, Instruct-CZ Centre, supported by MEYS CR (LM2023042, e-INFRA CZ (ID:90254)).
ER  -

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