Global SEC-PCP-SILAC mapping reveals protein complexes
mediating NF-κB activation in breast cancer

a 2022

Global SEC-PCP-SILAC mapping reveals protein complexes mediating NF-κB activation in breast cancer

LAPČÍK, Petr, Greg STACEY, David POTĚŠIL, Leonard FOSTER, Pavel BOUCHAL et. al.

Základní údaje

Originální název

Global SEC-PCP-SILAC mapping reveals protein complexes mediating NF-κB activation in breast cancer

Autoři

LAPČÍK, Petr, Greg STACEY, David POTĚŠIL, Leonard FOSTER a Pavel BOUCHAL

Vydání

In Book of Abstracts of 16th Central- and Eastern European Proteomic Conference, Praha 29.9.-1.10.2022, ThO-02, 2022

Další údaje

Jazyk

angličtina

Typ výsledku

Konferenční abstrakt

Obor

10608 Biochemistry and molecular biology

Stát vydavatele

Česká republika

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Organizační jednotka

Přírodovědecká fakulta

Příznaky

Mezinárodní význam

Změněno: 26. 11. 2022 23:36, Mgr. Petr Lapčík, Ph.D.

Anotace

V originále

NF-κB has essential role in immune response and is associated with lymph node metastasis of luminal A breast tumors [1]. Analysis of protein interactome and its changes in response to NF-κB modulation could uncover pro-metastatic mechanisms related to NF-κB. We apply metabolic isotope labeling SILAC, size exclusion chromatography (SEC) and protein correlation profiling (PCP) [2] to construct a network of interactome rearrangement in response to NF-κB modulation in MCF-7 breast cancer cells. We generated two co-fractionation datasets consisting of 80 fractions from SILAC-labeled and 80 fractions from label-free native MCF-7 lysates with inhibited or native NF-κB activity. LC-MS/MS analysis of SEC fractions using Orbitrap Lumos and Bruker Impact II mass spectrometers quantified 3308 and 5460 protein groups in SILAC and label-free datasets, respectively (FDR = 0.01). Interactome reconstruction using PrinCE [3] detected 7568 interactions among 1520 proteins. Co-elution of subunits of known complexes, such as ribosome, proteasome and MCM, was observed. Modulation of NF-κB was linked to interactome changes of proteins involved in immune response, cell cycle and DNA replication. NF-κB factor RELA interacted with proteins co-eluting with activators of NF-κB and these interactions were modulated by NF-κB inhibition. Our interaction network represents a complex insight into dynamics of MCF-7 protein interactome associated with NF-κB pathway and could serv e as a basis for future studies characterizing NF-κB in breast cancer.This work was supported by Ministry of Health of the Czech Republic (grant No. NU22-08-00230) and by the project National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union - Next Generation EU.

Návaznosti

LX22NPO5102, projekt VaV

Název: Národní ústav pro výzkum rakoviny (Akronym: NÚVR)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Národní ústav pro výzkum rakoviny, 5.1 EXCELES

NU22-08-00230, projekt VaV

Název: Proteogenomová klasifikace trojitě negativních nádorů prsu ve vztahu k prognóze a cílené terapii

Investor: Ministerstvo zdravotnictví ČR, Proteogenomová klasifikace trojitě negativních nádorů prsu ve vztahu k prognóze a cílené terapii, Podprogram 1 - standardní

Citovat

LAPČÍK, Petr, Greg STACEY, David POTĚŠIL, Leonard FOSTER a Pavel BOUCHAL. Global SEC-PCP-SILAC mapping reveals protein complexes mediating NF-κB activation in breast cancer. In In Book of Abstracts of 16th Central- and Eastern European Proteomic Conference, Praha 29.9.-1.10.2022, ThO-02. 2022.

@proceedings{2234357,
   author = {Lapčík, Petr and Stacey, Greg and Potěšil, David and Foster, Leonard and Bouchal, Pavel},
   booktitle = {In Book of Abstracts of 16th Central- and Eastern European Proteomic Conference, Praha 29.9.-1.10.2022, ThO-02},
   language = {eng},
   title = {Global SEC-PCP-SILAC mapping reveals protein complexes mediating NF-κB activation in breast cancer},
   url = {https://www.czechms.org/cz/events/cmsc/abstract.php?code=6210745},
   year = {2022}
}

TY  - CONF
ID  - 2234357
AU  - Lapčík, Petr - Stacey, Greg - Potěšil, David - Foster, Leonard - Bouchal, Pavel
PY  - 2022
TI  - Global SEC-PCP-SILAC mapping reveals protein complexes mediating NF-κB activation in breast cancer
UR  - https://www.czechms.org/cz/events/cmsc/abstract.php?code=6210745
N2  - NF-κB has essential role in immune response and is associated with lymph node metastasis of luminal A breast tumors [1]. Analysis of protein interactome and its changes in response to NF-κB modulation could uncover pro-metastatic mechanisms related to NF-κB. We apply metabolic isotope labeling SILAC, size exclusion chromatography (SEC) and protein correlation profiling (PCP) [2] to construct a network of interactome rearrangement in response to NF-κB modulation in MCF-7 breast cancer cells. We generated two co-fractionation datasets consisting of 80 fractions from SILAC-labeled and 80 fractions from label-free native MCF-7 lysates with inhibited or native NF-κB activity. LC-MS/MS analysis of SEC fractions using Orbitrap Lumos and Bruker Impact II mass spectrometers quantified 3308 and 5460 protein groups in SILAC and label-free datasets, respectively (FDR = 0.01). Interactome reconstruction using PrinCE [3] detected 7568 interactions among 1520 proteins. Co-elution of subunits of known complexes, such as ribosome, proteasome and MCM, was observed. Modulation of NF-κB was linked to interactome changes of proteins involved in immune response, cell cycle and DNA replication. NF-κB factor RELA interacted with proteins co-eluting with activators of NF-κB and these interactions were modulated by NF-κB inhibition. Our interaction network represents a complex insight into dynamics of MCF-7 protein interactome associated with NF-κB pathway and could serv e as a basis for future studies characterizing NF-κB in breast cancer.This work was supported by Ministry of Health of the Czech Republic (grant No. NU22-08-00230) and by the project National Institute for Cancer Research (Programme EXCELES, ID Project No. LX22NPO5102) - Funded by the European Union - Next Generation EU.
ER  -

LAPČÍK, Petr, Greg STACEY, David POTĚŠIL, Leonard FOSTER a Pavel BOUCHAL. Global SEC-PCP-SILAC mapping reveals protein complexes mediating NF-κB activation in breast cancer. In \textit{In Book of Abstracts of 16th Central- and Eastern European Proteomic Conference, Praha 29.9.-1.10.2022, ThO-02}. 2022.

Podrobný výpis o publikaci