ID | 61294 |
FullText URL | |
Author |
Nishioka, Keiji
Institute of Plant Science and Resources (IPSR), Okayama University
Kato, Yusuke
Institute of Plant Science and Resources (IPSR), Okayama University
Ozawa, Shin-ichiro
Institute of Plant Science and Resources (IPSR), Okayama University
Takahashi, Yuichiro
Research Institute for Interdisciplinary Science, Okayama University
ORCID
Kaken ID
publons
researchmap
Sakamoto, Wataru
Institute of Plant Science and Resources (IPSR), Okayama University
|
Abstract | Protein phosphorylation is a fundamental post-translational modification in all organisms. In photoautotrophic organisms, protein phosphorylation is essential for the fine-tuning of photosynthesis. The reversible phosphorylation of the photosystem II (PSII) core and the light-harvesting complex of PSII (LHCII) contribute to the regulation of photosynthetic activities. Besides the phosphorylation of these major proteins, recent phosphoproteomic analyses have revealed that several proteins are phosphorylated in the thylakoid membrane. In this study, we utilized the Phos-tag technology for a comprehensive assessment of protein phosphorylation in the thylakoid membrane of Arabidopsis. Phos-tag SDS-PAGE enables the mobility shift of phosphorylated proteins compared with their non-phosphorylated isoform, thus differentiating phosphorylated proteins from their non-phosphorylated isoforms. We extrapolated this technique to two-dimensional (2D) SDS-PAGE for detecting protein phosphorylation in the thylakoid membrane. Thylakoid proteins were separated in the first dimension by conventional SDS-PAGE and in the second dimension by Phos-tag SDS-PAGE. In addition to the isolation of major phosphorylated photosynthesis-related proteins, 2D Phos-tag SDS-PAGE enabled the detection of several minor phosphorylated proteins in the thylakoid membrane. The analysis of the thylakoid kinase mutants demonstrated that light-dependent protein phosphorylation was mainly restricted to the phosphorylation of the PSII core and LHCII proteins. Furthermore, we assessed the phosphorylation states of the structural domains of the thylakoid membrane, grana core, grana margin, and stroma lamella. Overall, these results demonstrated that Phos-tag SDS-PAGE is a useful biochemical tool for studying in vivo protein phosphorylation in the thylakoid membrane protein.
|
Keywords | Chloroplast
Phos-tag
Protein phosphorylation
Thylakoid membrane
STN7
STN8
|
Published Date | 2021-01
|
Publication Title |
Photosynthesis Research
|
Volume | volume147
|
Issue | issue1
|
Publisher | Springer
|
Start Page | 107
|
End Page | 124
|
ISSN | 0166-8595
|
NCID | AA00362200
|
Content Type |
Journal Article
|
language |
English
|
OAI-PMH Set |
岡山大学
|
Copyright Holders | © Authors.
|
File Version | publisher
|
PubMed ID | |
DOI | |
Web of Science KeyUT | |
Related Url | isVersionOf https://doi.org/10.1007/s11120-020-00803-1
|
License | http://creativecommons.org/licenses/by/4.0/
|
Funder Name |
Ministry of Education, Culture, Sports, Science and Technology
Japan Society for the Promotion of Science
|
助成番号 | 16H06554
17H03699
18K06290
|