Assessing the activation/inhibition of tyrosine kinase-related pathways with a newly developed platform.


Journal

Proteomics
ISSN: 1615-9861
Titre abrégé: Proteomics
Pays: Germany
ID NLM: 101092707

Informations de publication

Date de publication:
08 2021
Historique:
revised: 16 06 2021
received: 30 09 2020
accepted: 18 06 2021
pubmed: 22 6 2021
medline: 26 10 2021
entrez: 21 6 2021
Statut: ppublish

Résumé

The phosphorylation of cellular proteins plays a crucial role in the transduction of various signals from outside the cell into the nucleus. The signals are transduced by phosphorylation chain reactions within multiple pathways; however, determining which pathways are responsible for each defined signal has proven challenging. To estimate the activity of each pathway, we developed a phosphorylation array platform comprising a protein array with 1200 proteins belonging to 376 signalling pathways and an analytical method to estimate pathway activity based on the phosphorylation levels of proteins. The performance of our system was assessed by reconstructing kinase-substrate relationships, as well as by estimating pathway activity upon epidermal growth factor (EGF) stimulation and the pharmacological inhibition of epidermal growth factor receptor (EGFR). As a result, kinase-substrate relationships were reliably reconstructed based on the precise measurement of phosphorylation levels of constituent proteins on the array. Furthermore, the pathway activities associated with EGF stimulation and EGFR inhibition were successfully traced through the related pathways from the outer membrane to the nucleus along a time course. Thus, our phosphorylation array system can effectively assess the activity of specific signalling pathways that are perturbed by extracellular stimuli, such as various drugs.

Identifiants

pubmed: 34151541
doi: 10.1002/pmic.202000251
doi:

Substances chimiques

Tyrosine 42HK56048U
Epidermal Growth Factor 62229-50-9
Protein-Tyrosine Kinases EC 2.7.10.1

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

e2000251

Informations de copyright

© 2021 Wiley-VCH GmbH.

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Auteurs

Harumi Kagiwada (H)

Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Kotu-ku, Tokyo, Japan.

Takayuki Kiboku (T)

Socium Inc., Tokyo, Japan.

Hitomi Matsuo (H)

Socium Inc., Tokyo, Japan.

Masashi Kitazawa (M)

Socium Inc., Tokyo, Japan.

Kazuhiko Fukui (K)

Cellular and Molecular Biotechnology Research Institute, National Institute of Advanced Industrial Science and Technology, Kotu-ku, Tokyo, Japan.

Katsuhisa Horimoto (K)

Socium Inc., Tokyo, Japan.
Artificial Intelligence Research Center, National Institute of Advanced Industrial Science and Technology, Tokyo, Japan.

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