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

Investor logo

Warning

This publication doesn't include Faculty of Education. It includes Faculty of Science. Official publication website can be found on muni.cz.
Authors

LAPČÍK Petr STACEY Greg POTĚŠIL David FOSTER Leonard BOUCHAL Pavel

Year of publication 2022
Type Conference abstract
MU Faculty or unit

Faculty of Science

Citation
Description NF-?B 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-?B modulation could uncover pro-metastatic mechanisms related to NF-?B. 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-?B 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-?B 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-?B was linked to interactome changes of proteins involved in immune response, cell cycle and DNA replication. NF-?B factor RELA interacted with proteins co-eluting with activators of NF-?B and these interactions were modulated by NF-?B inhibition. Our interaction network represents a complex insight into dynamics of MCF-7 protein interactome associated with NF-?B pathway and could serv e as a basis for future studies characterizing NF-?B 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.
Related projects:

You are running an old browser version. We recommend updating your browser to its latest version.