Open Access

ARTICLE

Light-controlled phosphorylation in the TrkA-Y785 site by photosensitive UAAs activates the MAPK/ERK signaling pathway

by SHU ZHAO^1,*, SHIXIN YE²

1 School of Life Sciences, Nantong University, Nantong, 226019, China
2 Institute National de la Sante et de la Recherche Med icale (INSERM) U1195, Bicetre Hospital, Paris-Saclay University, Le Kremlin-Bicêtre, 94276, France

* Corresponding Author: Shu Zhao,

BIOCELL 2023, 47(6), 1377-1388. https://doi.org/10.32604/biocell.2023.023874

Received 19 May 2022; Accepted 06 September 2022; Issue published 19 May 2023

Abstract

Background: This paper aims to establish a light-controlled phosphorylation detection method at the Y785 site of tropomyosin receptor kinase A (TrkA) receptor in mammalian cells by using genetic code expansion technology and detecting the effects of optical activation of this site on the downstream MAPK/ERK pathway. The study is based on the current situation that the regulatory mechanism of TrkA phosphorylation has not been fully elucidated. Methods: Two photosensitive unnatural amino acids, p-azido-L-phenylalanine (AzF) and photo-caged tyrosine (ONB) were introduced into the TrkA-Y785 site by genetic code expansion technology and site-directed mutagenesis. Western blotting and laser confocal imaging were conducted to analyze the effects of this site on activating the MAPK/ERK pathway and nerve cell differentiation before and after photostimulation. Results: Our results supplemented the light-controlled results of the TrkA-Y785 site based on our previous research and verified that Y785 also makes important contributions in regulating the MAPK/ERK pathway. Conclusion: This study demonstrated the significant contributions of the TrkAY785 site in regulating the ERK pathway by precisely controlling the phosphorylation state of a single tyrosine site.

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Keywords

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