2016年12月
Probing the Lysine Proximal Microenvironments within Membrane Protein Complexes by Active Dimethyl Labeling and Mass Spectrometry
ANALYTICAL CHEMISTRY
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- 巻
- 88
- 号
- 24
- 開始ページ
- 12060
- 終了ページ
- 12065
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1021/acs.analchem.6b02502
- 出版者・発行元
- AMER CHEMICAL SOC
Positively charged lysines are crucial to maintaining the native structures of proteins and protein complexes by forming hydrogen bonds and electrostatic interactions with their proximal amino acid residues. However, it is still a challenge to develop an efficient method for probing the active proximal microenvironments of lysines without changing their biochemical/physical properties. Herein, we developed an active covalent labeling strategy combined with mass spectrometry to systematically probe the lysine proximal microenvironments within membrane protein complexes (similar to 700 kDa) with high throughput. Our labeling strategy has the advantages of high labeling efficiency and stability, preservation of the active charge states, as well as biological activity of the labeled proteins. In total, 121 lysines with different labeling levels were obtained for the photosystem II complexes from cyanobacteria, red algae, and spinach and provided important insights for understanding the conserved and nonconserved local structures of PSII complexes among evolutionarily divergent species that perform photosynthesis.
- リンク情報
- ID情報
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- DOI : 10.1021/acs.analchem.6b02502
- ISSN : 0003-2700
- eISSN : 1520-6882
- PubMed ID : 28193046
- Web of Science ID : WOS:000390621000015